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Cancer Cell Biology - Science topic

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Questions related to Cancer Cell Biology
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I have been working on Fluourescence in situ hybridization with HeLa cells. I hope to see noncoding RNA in HeLa cell. For negative control, I designed control set pretreated with RNase A(degrading all RNAs), and treated with specific probe. Before that, Samples were fixed with 4% paraformadehyde and permeablized. I wonder that RNase A is good working even at fixed sample?
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I fixed the cell with 4% PFA/1XPBS (pH7.4) for 15min and permeabilize the cell with 0.5% triton at RT for 10min. Next, we treat the cell with RNase If(NEB) overnight at 37.
To check wheter the RNA is removed, I stain the cell with SYBR gold. The nucleoli shows strong fluorescence.
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Logistics seeks both to address real-world problems and research questions and to unite terminological, conceptual and methodological research aspects. In the last 20 years, biological research has developed extremely powerful methods and tools for fundamental and applied research. There is furthermore an increasing desire to build integrative research platforms that combine interdisciplinary and multi-level data to the structural complexity of biological systems. How much attention do you give to logistics for your research plans? Do you think logistics supports research methods, innovation, quality, value, and impact as well as training and participation opportunities for students as well as for practitioners? If so how and if not why not.
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Andrew Lewis - You've made excellent points with clear analogies - thank you! I agree with your synopsis that "Transitioning biological understanding to this next level is certainly in large part a logistical problem" and that we must strive to the "coordination of manpower and computational power resources being put towards the pursuit of understanding things at this level."
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Just need to know if they grow (percentage of graft success, assuming one million cells in matrigel or similar conditions, orthotopic), and how long does it take, in order to plan some experiments.
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Following questions, does MDA-MB-415 xenografts easy to generate lung metastasis? How long approximately? Thanks!!
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What exactly is the role of HSP-90 in extracellular environment of the cell? I am wondering whether hsp90 is involved in the translocation of the client protein from outside to inside of the cell. If somebody is having some references please share with me. I am very curious about this molecule.
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My article about that just got accepted you can see it soon, including tests before, immediately and 2 hours after exercise
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Could someone kindly clarify why aerobic glycolysis is required for cancer cells? That is, if tumor cells used OXPHOS, would they be unable to grow and metastasize as well?
The most commonly cited benefit of aerobic glycolysis seems to be accelerated glucose uptake, but why is this required for long-term growth as opposed to short-term spurts? The timeline for cancer progression is normally months or years, not hours or days.
Moreover, despite consuming glucose faster, total ATP production from aerobic glycolysis is lower. That is, during the time OXPHOS consumes 1 glucose molecule, aerobic glycolysis may consume 13 glucose molecules -- yet this yields fewer ATP molecules.
If correct, total energy production inadequately explains the relationship between cancer cells and aerobic glycolysis.
Is the timing of ATP essential? Perhaps cancer cells need fewer shots of ATP molecules but at a higher frequency rather than wait for one large batch from OXPHOS?
Or what are the other benefits of aerobic glycolysis over OXPHOS?
Ultimately, why would cancer cells be less successful with OXPHOS?
Thanks in advance for your help.
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Aerobic glycolysis creates an acidic micro environment that compromises the TEJ of the normal native vasculature creating vasogenic edema which increases the glucose concentration in the tumor micro environment, favoring unregulated cellular proliferation. MREPT can detect vasogenic edema enabling minimally invasive ablation of cancer before cellular mass inhibits diffusion, and much before the formation of tumor vasculature can create “washout “.
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I'm preparing pancreatic cancer cells spheroids. Which concentration of collagenase can be used to dissociate them?
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Simona Mura Hi Simona, I realise this question was raised a few years ago now, but I am doing a very similar technique and wondered if you have an optimised protocol for this? Which vol/ conc did you end up using for your Pancreatic clusters? Thank you very much!
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This seminal paper by Paul Mischel rediscovered the relationship between extrachromosal DNA (ecDNA) and cancer:
1. How much would it cost to replicate the part of the experiment that generated subcutaneous tumors from seeds containing 200/2000/20000 cells of FACS-sorted EGFRvIII High/EGFRvIII Low subpopulations?
2. How much would it cost to sequence these tumor cells?
Thanks for your help!
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Shalini Sanyal Thanks for your answe.
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Hi, I am a beginner in the field of cancer genomics. I am reading gene expression profiling papers in which researchers classify the cancer samples into two groups based on expression of group of genes. for example "High group" "Low group" and do survival analysis, then they associate these groups with other molecular and clinical parameters for example serum B2M levels, serum creatinine levels for 17p del, trisomy of 3. Some researchers classify the cancer samples into 10 groups. Now if I am proposing a cancer classification schemes and presenting a survival model based on 2 groups or 10 groups, How should I assess the predictive power of my proposed classification model and simultaneously how do i compare predictive power of mine with other survival models? Thanks you in advance.
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The survAUC R package provides a number of ways to compare models link: https://stats.stackexchange.com/questions/181634/how-to-compare-predictive-power-of-survival-models
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Hi There,
I have attempted to perform a double-thymidine block on my cells using a 24h first inc.-9h release-16 second inc. and 16-9-16 incubation periods in FUCCI DLD-1 cells. I haven't seen any synchronization. Does anyone have any recommendations or tips?
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In my opinion you need to release the cells from thymidine to see the effect of arrested population especially with fucci cells. The thymidine double blocked cells will give you the similar profile as asynchronized cells but when you release the cells into fresh medium after the second block you can get the enriched population of cells in S phase.
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I tried staining MCF7 and MB231 for Vimentin and Cytokeratin antibodies, and surprisingly, I saw both cell lines showed both markers. According to the literature, MB231 is a mesenchymal type cell and MCF7 is luminal, so shouldn't it be that MB231 shows more cells positive for Vimentin as compared to MCF7?
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Hi Rafał Watrowski . Adity Pore was just asking why markers for the hypothetical EMT do not make sense in the 2 cell lines she is looking at.
I agree. The EMT is just a hand waving explanation of how epithelial cells, that are normally sedentary, suddenly become motile and invasive when they are cancerous.
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I am already aware that HepG2 and Hep3B are both HCC cell lines, but due to site-permission issues I am not able to access most literature. I need an etiological as well as characteristic differences between the two. Can anyone help?
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We just identified that Hep3B has p53-FXR2 fusion due to 76 kb focal deletion within chromosome-17 in this paper
Mutant p53s and chromosome 19 microRNA cluster overexpression regulate cancer testis antigen expression and cellular transformation in hepatocellular carcinoma | Scientific Reports (nature.com)
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My research is to analyse the estrogenic activity of possible substances that migrate from plastics using the E-screen test based on the ability of MCF-7 cells to proliferate in the presence of oestrogens. However, so far I have not been able to validate the assay since I have failed to see significant differences between the effects caused by my positive control (17β oestradiol at almost any concentration) and my negative control. There is one paper that suggest that different "sublines" of MCF 7 cells show different sensitivities to oestrogen. So, I was wondering if you have been able to see differences between the proliferation in the presence of E2 and in presence of the negative control. and where did you get your mother stock of MCF-7
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It may be the presence or absense but above all I want to make sure it is like this. The rest could be sensitivity
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Does anyone have an IHC protocol for detection of a his-tagged protein in mouse tumor tissue (cryo, not paraffin)?
I'm trying to stain for a protein that contains a 6His tag and accumulates in tumor tissue (human tumor xenograft grown in mice). I used the M.O.M. blocking kit from Vector Labs to block endogenous mouse IgG, then incubated with the mouse penta-his primary antibody. The secondary antibody used was the Vector Labs Impress HRP-anti-mouse antibody. The problem is I get really high background signal, even in the negative control (no primary antibody). It seems the only species of penta his antibody is mouse derived, so maybe someone knows a source of rabbit, goat, or donkey penta his antibody?
Thanks,
Jason
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I am curious which goat polyclonal anti-6his antibody did you end up in using from abcam? Thanks!
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Hi I am trying to see which Rab5 isoform would be ideal to track E-Cadherin exocytic vesicles. Does anyone have experience with this?
Thanks,
Mukhtar
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A study by Peeters et al. (2017) suggests that sugar traps cancer in a 'vicious cycle' which make it more aggressive and harder to treat (1). On the question-and-answer site Quora, Ray Schilling, MD, concludes: "there is a connection between the consumption of sugar and starchy foods and various cancers in man. Animal experiments are useful in suggesting these connections, but many clinical trials including the Women’s Health Initiative have shown that these findings are also true in humans. It is insulin resistance due to sugar and starch overconsumption that is causing cancer" (2).
References
1. Peeters K, Van Leemputte F, Fischer B, Bonini BM, Quezada H, Tsytlonok M, Haesen D, Vanthienen W, Bernardes N, Gonzalez-Blas CB, Janssens V, Tompa P, Versées W, Thevelein JM. Fructose-1,6-bisphosphate couples glycolytic flux to activation of Ras. Nat Commun 2017; 8: 922. doi: 10.1038/s41467-017-01019-z. https://www.nature.com/articles/s41467-017-01019-z.pdf
2. Schilling R. Why isn't sugar portrayed as bad like cigarettes? https://www.quora.com/Why-isnt-sugar-portrayed-as-bad-like-cigarettes
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The fact that cancer cells use more sugar than non-cancer cells, known as the Warburg effect, appears to form the basis of the current question. Nevertheless, it is not yet known if this effect is merely a symptom of cancer or whether it is the cause of cells becoming cancerous. Evidence indicating the way cancer cells metabolize sugar is not conclusive, either. Considering refined or added sugar, the bulk of the available evidence indicates that sugar is not a carcinogen. There is no evidence that eating sugar causes cancer or speeds up its growth. There is also no evidence that eliminating dietary sugar causes cancer to shrink or disappear. However, it is known that eating excess sugar, especially added sugars, can contribute to many chronic diseases such as obesity and diabetes, which are potential risk factors for several types of cancer.
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Hi everyone,
I want to use the amplex red assay to quantify H2O2 levels in different cellular clones. Many protocols I found online or previous answers on RG contradict each other.
The major debate is about performing the assay:
- on cultured cells directly
or
- on culture media removed from cell cultures
What's best in your opinion ?
Someone also said that phenol red could interfere with the assay.
Thanks for the help you can provide !
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Hi everybody,
I just came across this publication
stating that amplex red can be converted into the fluorescent resorufin compound without requiring H2O2. The authors propose to conteract this effect by aPMSF treatment.
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Is there any way in which we can check whether the DNA inside the cell is negatively or positively supercoiled?
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Techniques to study DNA supercoiling include agarose gel electrophoresis and psoralen-based molecular probes. Using these approaches, unrestrained DNA supercoiling has been identified and mapped in eukaryotic genomes.
Good luck
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I am working on mechanism study of some anticancer compounds using A549 and H460 NSCLC cell lines. I want to check if my compounds are non-toxic to normal cell or not. Some researcher use human normal lung fibroblasts HEL-299 and MRC-5 cells while others use normal cell lines that don't belong to lung tissue. I want to use only normal lung cell, not from other tissue. Can anyone suggest me which one is the best normal lung cell line model?
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Hi there, may I know if the results are reliable and valid when we are to compare between two cell lines composed of different cell types? For example, A549, the epithelial cells, and MRC-5, the fibroblasts? Anyways, I have seen a lot of researchers using MRC-5 as control cell lines when they are working on A549 cell lines.
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I know rapamycin is generally used as an immunosuppresive agent in the treatment of organ transplantation (kidney transplantation) and works through inhibiting the mTORC1 complex. Additionally, due to the inhibition of mTORC1, rapamycin also produce some anticancer effect. So, my question is, if the immunity get suppressed because of rapamycin effect how it will provide anti-cancer effect? As cancer cells could proliferate more due to the low immunity of the body. Looking forward to hearing from the experts. Thanks in advance.
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Multiple anti-cancer effects. Some examples in the attached files.
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Hi everyone,
Does anyone know a really working mitochondrial isolation protocol? I need a quick separation method for a large number of cells. A quick protocol is needed because the substances that I am planning to determine are unstable.
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Human/rat/mouse tissue was minced and homogenized in isolation buffer [320 mM sucrose, 5 mM Tris (hydroxymethyl) methylaminoethane sulfonic acid (TES), 1 mM EGTA, pH 7.2]. The homogenate was then centrifuged at 1000 g for 5 min at 4 °C. The supernatant was collected and the pellet was resuspended in isolation buffer followed by second round of homogenization and centrifugation as mentioned earlier. The supernatants from both the steps were then pooled and centrifuged at 8,500 g for 10 min at 4°C. The crude mitochondrial fraction obtained in the pellet was resuspended in minimum volume of isolation buffer (~200 µl), overlaid on 6 % (w/v) ficoll solution, and centrifuged at 75,000 g for 30 min at 4°C, to remove the myelin content. The pellet was then resuspended in ~150 µl reconstitution buffer (250 mM sucrose, 10 mM TES, pH 7.2) and stored at -80 °C.
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Hi all,
I have kept cells in -80 for several months, can I transfer them to liquid nitrogen now after all this time? I am afraid they would not survive.. I appreciate your input
Thanks!
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It should be ok. You can keep & use later on. - 80 degree C, commonly use for short time use e.g 1-3 months. LN for longer periods of time. Best of luck!
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  • I trying to RNA isolate from mice blood which technique is best?
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Generally, horizontal gel electrophoresis is an ideal choice for DNA and RNA separation, while vertical systems are ideal for proteins.
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Aneuploidy, an imbalanced complement of chromosomes, was identified as a distinct feature of cancer cells more than a century ago. Indeed, ~90% of solid tumors are aneuploidy. The random gain and loss of complete chromosomes has many favorable properties as a mechanism for phenotypic switching, both in the context of a preemptive bet-hedging strategy as well as a rapid means for coping with unfamiliar environmental stresses. Aneuploidy might be a useful bet-hedging strategy for responding to environmental perturbations that are not frequently encountered. As aneuploidy perturbs hundreds of genes simultaneously, it has the potential to affect larger numbers of genes and processes per event relative to a spontaneous mutation. Importantly, unlike a spontaneous mutation, aneuploidy and any associated growth defects can also revert at a high frequency (cancer reversion).
So, aneuploidy may be the proof of “cancer is adaptation mechanism”
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Dear Andrzej and dear Mezut,
Polyploidy usually results in aneuploidy but it (polyploidy) has the means to counteract aneuploidy (through some variant of somatic meiosis). In turn, aneuploidy may also have its contribution to adaptation by clonal selection, even if transient. Likely, it is a balanced process resolving at the population level. Our review of meiotic genes in cancer is just accepted, where we tried to update this conundrum. After I get the proofs, I shall attach it here.
Katrina
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I would like to label Human Serum Albumin for my experiments.
My supervisor suggested NHS and 6-Aminofluorescein as working agents.
Can anybody please suggest a working protocol?
Any suggestions on storing? Can I freeze the labelled protein in a solution?
If you have any other suggestions I would be very grateful!
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Obviously numerous examples of how you can perform the labelling can be found. It depends on whether you use the label(s) separately or as an already prepared ready-to-use sample like in:
In essence it comes down to the use of NHS in order to enable the fluorescein to get attached to your protein. Once the labelling is finished you need to rid of the excess of NHS and 6-aminofluorescein. In:
you see that they use dialysis to remove the excess of NHS and 6-aminofluorescein. In:
Bidmanova, S., Chaloupkova, R., Damborsky, J., & Prokop, Z. (2010). Development of an enzymatic fiber-optic biosensor for detection of halogenated hydrocarbons. Analytical and bioanalytical chemistry, 398(5), 1891-1898.
the conjugates (CF–BSA) were separated from unreacted dye by size-exclusion chromatography.
Your question about storage. Indeed you can make one stock solution (let say 20 mL) and make smaller portions (let say 0.5 mL) and freeze them. Once you take a portion out of the freezer you better use it one time and better not freeze it back again (to be one safe side).
Best regards.
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In my experiment, I need to use ascorbic acid to treat cancer cells. As we all know ascorbic acid is very unstable in aqueous solution because of its oxidizing behavior, can anyone please tell me if I want to do treatment with ascorbic acid to cell lines then how could I prepare it? Should I adjust ascorbic acid to PH 7?
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Hi,
I see you already got some answers and mine won't be very different. Ascorbic acid is very reactive and easily oxidised. You could freeze it if you put some nitrogen gas through it first. However that might be more work than preparing it each time fresh.
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I want to know the experimental and technical differences between the breast cancer cell lines SUM159 and MDA-MB-231 cells. I wonder, do they have efficiency in sphere-formation( Stemness).
please help me with this.
Thank you
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MDA-MB-231 and SUM159 can both be cultured as oncospheres in Onco2 medium (HUMEC + Pen/Strep). But the SUM159 create a layer around that spheres that can be visualized by microscopy. This layer makes it hard to dissosiate all the cells properly for example FACS analysis, but protocols such as western blot with the lysis buffer will solve this problem. I use both cells for culturing oncospheres myself
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Do you know any normal mouse lung cell lines other than MLE 12 (which transformes Tumorogenic)?
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Hi Robert,
I found that Wi38 is a human lung fibroblast line, not from mouse. It is mentioned in the article you attached and also in the Sigma Aldrich website from where I wanted to get it.
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The given IC-50 concentration in the data sheet of inhibitor 10 micro molars, to look at the inhibition of IL-8. The concentrations above the IC-50 seem to be giving considerable results for eg. the concentration of 20 micro molars. The methodology used for the detection is IL-8 Indirect ELISA. Kindly suggest. 
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Cancer drug discovery
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The suggestions offered here are very helpful. I have the same issue, and would also appreciate some journal articles to cite if anyone knows of any. It seems that this phenomenon has happened to a few others. Many of the answers above can apply to our drug- the discovery that the manufacturer supplied an incorrect structural component and the chemist combined correct and incorrect components to generate the final compound could explain the inhibition to targeted cell, but promotion to a different non-expressing cell. But the separated compounds still exhibit toxicity at low concentration and slight promotion at high concentration, though selectivity for target is not happening with the incorrect product. It does seem that metabolism and a temporary cell cycle arrest may be playing roles. Or it could be that binding becomes non-specific at high concentrations so that the drug binds to unintended targets. Or it could be due to bio availability if the drug's solubility is dependent on pH and concentration.
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Hello Everyone,
I have some questions related to a drug called Cobalt 2 chloride hexahydrate.
I am currently preparing an experiment related to Hypoxia. I bought a drug called " Cobalt 2 chloride hexahydrate" (from Sigma Aldrich) with the molecular weight of 237.9 g/mol and the total weight is 5g.
I want to know what is the best stock concentration should we make for this drug? Plus, how much H20 should we add to the drug ? and the optimal condition to store this stock concentration?
Furthermore, can you recommend me the best final concentration of this drug to mimic the Hypoxia environment? any ideas?
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Dear Sv Mg, you're very welcome. Normally, cobalt(II) chloride itself is rather stable under normal conditions. However, you can't do anything wrong when ypu store the stock solution at 4 °C in the refrigerator.
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Hello,
I want to work with Resveratrol against some cancer cells. I want to know your experience about its Non-toxic dose by using MTT, its solubilty and some other things. Please let me know if someone is working with it. Thanks.
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Resveratrol downregulates inflammatory pathway activated by lymphotoxin α (TNF-β) in articular chondrocytes: Comparison with TNF-α. Buhrmann C, Popper B, Aggarwal BB, Shakibaei M. PLoS One. 2017 Nov 2;12(11):e0186993.
Resveratrol Regulates Colorectal Cancer Cell Invasion by Modulation of Focal Adhesion Molecules. Buhrmann C, Shayan P, Goel A, Shakibaei M.
Nutrients. 2017 Sep 27;9(10).
Sirt1 Is Required for Resveratrol-Mediated Chemopreventive Effects in Colorectal Cancer Cells. Buhrmann C, Shayan P, Popper B, Goel A, Shakibaei M.
Nutrients. 2016 Mar 5;8(3):145.
Resveratrol induces chemosensitization to 5-fluorouracil through up-regulation of intercellular junctions, Epithelial-to-mesenchymal transition and apoptosis in colorectal cancer. Buhrmann C, Shayan P, Kraehe P, Popper B, Goel A, Shakibaei M. Biochem Pharmacol. 2015 Nov 1;98(1):51-68.
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We have mice injected w/ specific cancer cell lines. The cancer cell lines failed to pick up the luciferase bioluminescent tag. We have a rubric of categories to measure the mice, but I was curious to see if there were any other variables that we could measure that would give proxy measurement for changes in tumor size.
thank you.
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We inject the tumor cells in the flank and we measure the palpable tumor after 7 days using a caliper as said previously. We also use bioluminescence imaging to quantitate the data.
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I would like to study development mechanism of colon cancer, could anyone give some advice about the advantage or feature of each of FHC and CCD 841 CoN?
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Debasish Kumar Dey
Thank you so much for your information!
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I wanted to know if we can store frozen cells in -80 C refrigerator. These cells are stored in liquid nitrogen tank (-200 C) as of now. But as liquid nitrogen vaporizes, we need to keep refilling the tank every two months. Since, the university is going to be under lockdown due to COVID-19, we will not be allowed to enter labs or be able to order new liquid nitrogen. So will it be feasible to store these frozen cells at -80 C for the coming 3-4 months?
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Hi Adity,
I think you can keep your cells in -80 for several years. However I strongly suggest that after the Covid-19 situation, cultivate your cells again and prepare your nitrogen source from fresh culture. Do not put them directly into Nitrogen Tank.At least make an experiment first. For example, put some cryovials into liquid nitrogen and after couple of days, cultivate them. If they are alive you can transfer your cells into liq. nitrogen tank too.
The first concern for me is contamination.
Second, depen on my experiences, I realized that cells dye if they stay first too long in -80 and consequenlty transfered to nitrogen. With too long, I mean more than 2 months.
Cells are normaly alive -80 for years. I agree with that. However, if you put these cells from -80 after 3 months into liquid nitrogen, unfortunately cells may dye. That occured to us several times. That's why we never keep our cells more than 3 weeks in -80 after crypreservation protocol. If we forget them in -80, we use them but never put them into nitrogen tank again.
I hope your cells will be Ok.
Good luck!
Cigdem
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Dear friends and colleagues,
there is a very important resource on the web that is "Atlas of Genetics and Cytogenetics in Oncology and Haematology". The Atlas is a peer reviewed on-line journal / encyclopedia / database established in 1997 that collects data about cancer genes, solid tumors, leukaemia and other resources. It is in open free access for readers and there are no fees for the authors.
I'm searching volunteers and collaborators that would to write with me reviews about single genes involved in cancer.
I have published several papers so far and I believe that Atlas is a very valuable tool for us researchers, scientists and scholars.
I invite you to visit the Atlas website at http://atlasgeneticsoncology.org/index.html for more information. Here is my latest article published http://atlasgeneticsoncology.org//Genes/GC_EEF1B2.html to show you my working method. Anyone wishing to collaborate with me to write a specific review on a gene can tell me as well.
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I have research and interests in medical genetics on a less laboratory research results level. I rely on the scientific research in Atlas of Genetics and Cytogenetics in Oncology and Haematology nonetheless. Congratulations on your ambitious and important research project! Best regards.
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Oncology is a branch of medicine that deals with the prevention, diagnosis, and treatment of cancer. A medical professional who practices oncology is an oncologist. The name's etymological origin is the Greek word ὄγκος (óngkos), meaning 1. "burden, volume, mass" and 2. "barb", and the Greek word λόγος (logos), meaning "study".
Cancer survival has improved due to three main components: improved prevention efforts to reduce exposure to risk factors (e.g., tobacco smoking and alcohol consumption), improved screening of several cancers (allowing for earlier diagnosis), and improvements in treatment.
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Lauren Pecorino
Molecular Biology of Cancer: Mechanisms, Targets, and Therapeutics
3rd Edición
ISBN-13: 978-0199577170, ISBN-10: 019957717X
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Metformin's anti-cancerous properties have been demonstrated in various cancer cells in vitro, such as lung, pancreatic, colon, ovarian, breast, prostate, renal cancer cells, melanoma, and even in acute lymphoblastic leukemia cells. 
It was suggested that the tumoral  microenvironment is associated with long-term outcome in primary and metastatic tumors.Metformin reduces inflammatory microenvironment Is regulated microenvironment with metformin reprogramme malignant cancer Cells toward a benign phenotype.
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Epigenetic therapy inhibits metastases by disrupting premetastatic niches
  • Zhihao Lu,
  • Jianling Zou,
  • […]
  • Malcolm V. Brock
Nature volume 579, pages284–290(2020
Abstract
Cancer recurrence after surgery remains an unresolved clinical problem1,2,3. Myeloid cells derived from bone marrow contribute to the formation of the premetastatic microenvironment, which is required for disseminating tumour cells to engraft distant sites4,5,6. There are currently no effective interventions that prevent the formation of the premetastatic microenvironment6,7. Here we show that, after surgical removal of primary lung, breast and oesophageal cancers, low-dose adjuvant epigenetic therapy disrupts the premetastatic microenvironment and inhibits both the formation and growth of lung metastases through its selective effect on myeloid-derived suppressor cells (MDSCs). In mouse models of pulmonary metastases, MDSCs are key factors in the formation of the premetastatic microenvironment after resection of primary tumours. Adjuvant epigenetic therapy that uses low-dose DNA methyltransferase and histone deacetylase inhibitors, 5-azacytidine and entinostat, disrupts the premetastatic niche by inhibiting the trafficking of MDSCs through the downregulation of CCR2 and CXCR2, and by promoting MDSC differentiation into a more-interstitial macrophage-like phenotype. A decreased accumulation of MDSCs in the premetastatic lung produces longer periods of disease-free survival and increased overall survival, compared with chemotherapy. Our data demonstrate that, even after removal of the primary tumour, MDSCs contribute to the development of premetastatic niches and settlement of residual tumour cells. A combination of low-dose adjuvant epigenetic modifiers that disrupts this premetastatic microenvironment and inhibits metastases may permit an adjuvant approach to cancer therapy.
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I guess I'm not understanding the concept well. If I treated cells (5000/well) with a range of doses from 6nM to 92nM (and this is an MTT assay and I've used Prism8), why is the IC50 only .9nM? I guess I'm not reconciling what I'm putting into the assay and what prism spits out. Are the values from Prism in the same units as my initial concentration? I also log transformed the values so Idk if that has anything to do with it.
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Hello Luna,
Maybe it would help if you posted your curve.
If you inserted your data, i.e. concentrations and corresponding effects into prism it will try to "fit" your data points on a curve and calculate an IC50 value. Without knowing more details of your experiment I suspect:
If the IC50 ends up to be 0.9nM while your lowest conc. was 6nM, it would mean that you drastically overdosed your cells (as mentioned above) and 6nM gets viability/survival way below 50%. So the algorithm will draw a line between x=0/y=100% (untreated) and x=6nM/y=(possibly)10%. Following that line, if you reach y=50% your x will be 0.9nM. (Which is, of course, highly inaccurate!)
Bottom line: (As also mentioned above) you should repeat the experiment with lower concentration range.
Hope that helps
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In the body, when one develops a tumor, the microenvironment is very important. How do the cell cultures of cancer cells grow in the flasks without that micro-environment and only with the media and the serum?
Any suggested readings?
Thanks.
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Very good question. The real microenvironment of cancer cells in vivo is very harsh: acidic, hypoxic, poor in nutrients, high in CO2.
They adapt to all these and through clonal selection in a Darwinian fashion they live where other cells cannot.
When you put then in a flask everything is better for these die hard cells. Life is easy in the flask.
These are reasons why what you achieve in an experiment in the flask is not reproducible in vivo.
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Is it because we are trying to target these cells before they have a chance to divide??? I guess I don't understand the logic behind this.
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"Low" confluence (30-50%) should be used for evaluating cytostatic effects. In other words, how effectively a drug will be able to stop actively proliferating cells from dividing (cell cycle arrest). We use this confluence so that the cells will have space to grow and divide. For example: if you want to test the cytostatic effect of a drug and you seed cells at a confluence of 70%. Chances are that by tomorrow (exactly prior drug treatment), you will have a confluence close to or equal to 100%. You won't be able to see differences between treatment groups since the drug will only affect proliferation, but they have already reached confluence (they cant proliferate further from the physical limit of the monolayer and the well of the plate). The lower range of confluence will be determined by the sensitivity of the assay used to detect the cells and the higher range of confluence by the proliferation rate of your cells. (e.g. MDAMB231 cells are highly proliferative, i will opt for seeding them at 20% confluence, SKBR3 proliferate slowly, i will seed them at 50% confluence for addressing a cytostatic effect of a drug)
High confluence (70-90%) for a cytotoxic effect. In other words, how effectively a compound will be able to induce cell death in your cell population. In this case, initial confluence is more flexible, but it will always start from a determined amount of cells and after the treatment, you will have the same amount of cells (if the treatment has no cytotoxic effect) or less.
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I'm focusing on the Ras/Raf/MapK/Erk pathway and I'm expecting to see both when I treat with a MEK inhibitor in pancreatic cell cancer line. While I dramatically reduced levels of pERK, there's VERY little cell death. Does this mean that apoptotic pathways aren't necessarily rescued in this case or it is a technical issue where I should leave my cells in the treatment for longer than my current timepoint?
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6 could be indeed quite a short time to see apoptosis. I would also suggest you to start with a time-lapse experiment rather than an end point experiment. It is sufficient to have a nuclear marker to recognize apoptosis, or even in white-field illumination. With a time lapse experiment you can immediately realize when apoptosis occurs and the later to do an end-point experiment with the time that you learned from time-lapse.
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I am trying to grow cancer cells and do MTT assay for drug screening. Are there any alternatives to FBS? I know that FBS and FCS are widely used for cell culturing, but I am looking for other options.
I found one online and got it ordered:
But the cells are not growing properly in this.
Any suggestions in this regard?
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You can try thermofisher product for serum free media
https://www.thermofisher.com/in/en/home/life-science/cell-culture/mammalian-cell-culture/serum-free-media. As Adam Shapiro has mentioned, you have to slowly adapt the cultures to serum free conditions. You can also try using DMEM/Ham's F-12 (50:50) supplemented with growth factors and hormones.
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Is there any portal or application that accepts Seurat objects as input and lets me see genes’ expression across the clusters? Of course, my lab’s service provider can help using Seurat itself, but I need to come to them several times when I have a request. I am learning Seurat, but it would be nice to have a tool that supports it upfront without command lines.
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Hi Jaliu, you can try out BioTuring Browser (www.bioturing.com/bbrowser). The software takes in Seurat and Scanpy objects for visualization (keeping the same t-SNE or UMAP coordinates you have created using such tools) and extra analyses like marker finding, composition and differential expression analysis.
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I am looking for a simple and low-cost method of monitoring the cholesterol content in the cell membranes. My initial idea was to stain them with cholera toxin (anti-GM1 ganglioside) to get information about lipid rafts, so indirectly about the cholesterol content.
Do you have any better ideas?
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You have different options. The presence of cholesterol in the membrane gives rigidity, while the outflow of cholesterol causes membrane fluidity. Thus, the measurement of membrane fluidity can be an indirect measure of cholesterol levels in your cells. Another option is the use of fillipin. I leave you some links that I hope will serve you.
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i want to knoe in details hoe to calcule the cut off value for roc curve and hoe to make this rock curve and the calculation formula for sensistivity and specificity
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Thanks for valuable answers
Also one of the best software is Medcalc
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Hello everyone,
I have two questions, please shed some light on this. I am new to in vivo studies and some may find my questions are trivial. TIA.
My first question is,
I wanted to develop the MCF7 mouse xenografts. I did a search and came to know that the injection of estradiol is recommended to develop tumours prior to injecting the MCF7 cells. Also, I have seen some papers without estradiol. Can you tell me the role of estradiol in tumour formation? Can the mice form tumour without estradiol?
Lastly,
Once the tumour reaches a definite volume, I wanted to inject my compound of interest. Here is the question, which is a simple way to inject? I have found the following ways related to my studies.
1. Trochar
2. Microosmotic pump
3. Intraperitoneal
4. Oral gavage
Please recommend me a simple way to introduce my compound to the mice. If you have a better option also welcome.
Many thanks in advance,
Arun.
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Thank you, everyone, for your valuable suggestions. I have seen in a few research papers that people developed tumours without using estradiol supplements. Is it possible to do so? TIA.
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autophagic activity suppression by Rubicon is a signature of aging.
how many time it takes in the suppression of autophagic activity by Rubicon?
When Rubicon functions in neurons?
How Rubicon functions in neurons?
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Autophagy, an evolutionaily conserved cytoplasmic degradation system, has been implicated as a convergent mechanism in various longevity pathways. Autophagic activity decreases with age in several organisms . Research has shown that the expression of Rubicon, a negative regulator of autophagy, increases in earth worm , fly and mouse tissues at transcript and/or protein levels, suggesting that an age - dependent increase Rubicon impairs over time, and therby curtails animal health-span.. Consistent with this idea, knockdown of Rubicon extends worm and fly lifespan and ameliorates several age - associated phenotype. For more information consult nature.com
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I'm looking for information of AgNPs in particular
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I am a bit confused with this discussion. Personally, I don't think the size of the nanoparticle is the critical issue. The important thing is what you do with them- how you tag the chemotherapeutic agent, how you direct the functionalised nanoparticle to the cancer tissue, the mechanics of drug release, the mode of cell kill (chemotherapy or thermal or something else) and the fate of the nanoparticle after it has done its job. Cancer therapy is multi-facetted and focusing on one aspect of the therapy (size of the particle) may be ill advised..
Thank you.
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I want to know whether tumor cells share their information by time passing. I were wondering if anybody could answer my question and introduce some good resources in this regard?
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The answer is "Yes". Please see the following PDF attachments.
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I like to know the possible markers of metabolism in cancer cells such as in monitoring glycolytic, extracellular acidification and oxygen consumption rates, effected by treatment with an extract.
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It depends if you are fixing the cells for cellular markers or keeping them viable for live cell imaging markers. Also, whether you want to measure expression induction markers (mRNA expression), or on-going cellular processes (proteins, subcellular organelle activity like mitochondria and lysosomes).
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I have overall 2 years of Research experience including my Master thesis project. I have co authored 2 publication in international journal. One poster in international conference. I have cleared IELTS exam with 6.5 band in 2018.
I am looking for PhD admission.
My area of expertise and interest are: oncology, reproductive biology and developmental biology.
any help and guidance, I will highly appreciate.
Thanks in advance.
Kindly feel free to comment or inbox me.
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Dear Tariq,
One way: First step will be to contact some potential Principal investigators in USA and Europe, Australia, nzl (Canada does not have much reproductive biology but can try). If he agrees he will tell you the procedure to apply thru his/her University
Second way: In india, there are commonwealth scholarships which are for doing PhD in Commonwealth countries. Once you apply that scholarship, simultaneously get some support letter from potential PI a letter (they will be happy to give you since they are not giving you any money) that will help during interview process
Thirdway: See adds in Researchgate and Linkedin
Fourthway: Buy a book from your bookseller "Study Abroad"
Fifthway: Visit 5/6 embassies personally with your resume and papers (European mainly), they may have some exchange programs and talk to educational attache if he has any scholarship to do PhD for you
Good luck
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I am studying gene expression profile through analyses of proteins expressed upon treatment of cancer stem cell line with a novel herbal substance (in the extract) to predict gene function in the mechanism of reaction. I would, therefore, need to know the best method to determine/screen proteins expressed solely consequent upon this treatment in achieving this considering;
1. Model of the study (cancer stem cell line)
2. Testing molecules (drug candidate from herb)
3. Analyses involved.
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Hello
The SILAC/ACT followed by LC-MS/MS it's a good suggestion.
Good Luck
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I am currently looking for academic professional that can accept my studentship for PhD program in any of the following research area:
1. Antimalarial or anticancer novel compound synthesis and biochemical evaluation.
2. Structural biology of uncharacterized enzyme target for malarial or cancer therapy.
3. And other related research topics.
These studies will involve the use of bioinformatics/computational and biochemical/biophysical techniques.
Country of interest: Brazil, USA, Germany and other available countries.
Kindly let me know your willingness
Kind regards.
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Do you have your own funds?
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Hi!
I will be graduating soon with an MSc in cancer pharmacology from the UK. I want to know what job titles/description should I apply to in Singapore. my practical experience is mainly in cell culture. I also have a BSc in Pharmacy.
Any help is appreciated.
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I will suggest you to go for higher studies i.e. PhD in Oncology. or R&D section in Pharma and Biotech industries focus on Cancer biology.
Note: Go for IELTS/TOEFL along with GRE. It would help a lot. Atleast English test minimum.
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I am attempting to stain cells in my colony forming assay and would like to find a protocol, specificaly one that does not involve formaldehyde or buffered formalin.
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result.
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Im co-culturing the caco2, Ht29-MTX cell lines (ratio 1:10) on a silicon porous membrane by the pore sizes of 6um (center-center 50um). Actually I dont use the SERUM-FREE media and I do not know why do they use it in litreture for caco2 cell lines cultivate.
The result is that I see the cells in the holes but they are almost stoped to pass to the other side. And on the top surface of membrane, there are only clusters of cells instead of the cell monolayer.
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Try using Serum-Free medium.......
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what is the simplest ways to decontaminate an infected incubator and laminar from fungal contamination ?
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PFA = Paraformaldehyde See above. I would use all three in succession. Start with PFA, then Bleach, then EtOH. You may also want to consider Hydrogen peroxide instead of Bleach.
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Can somebody please tell me a most apt media composition for isolated fibroblasts which would mimic a maximum limit of its in Vivo microenvironment ? Thanks
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Thank you very much ...
Steingrimur Stefansson yes these are primary fibroblasts and thanks your recommendation would be great help....🙂
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I want to use data analysis to figure out how to predict the factors affecting the incidence of brain and bone cancer, but my challenge is the reliability of the data provided on the kaggle.com website. Is it possible to trust the data?
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I have several microarray gene expression datasets, but they use different naming systems for the probes. What are some simple ways to convert all the probes to the same type (e.g. Affymetrix)? Thanks.
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Hey there,
Here you can easily convert different naming.
Hope this helps.
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Hi, I am currently design a assay for testing the cytotoxicity of primary T cells against tumor cell lines. Here is the design:
Cryo-preserved PBMCs were retrieved and cultured with 9ug/mL PHA, 20%FBS RPMI1640 medium for 48 hours. Then the PBMCs were stained with CD3 antibody and were sorted for CD3+ T cells. The adherent tumor cell lines A549 were seeded the day before add T cells in 96 well plate (3000 cells/well). Tumor cells were stained with LIVE/DEAD® Viability/Cytotoxicity Kit (Themo Fisher: L3224). After T cell purification and tumor cell staining, we add T cells to tumor cells in 200 ul medium. Finally, the cytotoxicity were observed in In-Cell-Analyzer 2200 (GE) for 6 hours by real-time imaging and count for red signal of dead tumor cells. The RNA of T cells were collected after recording of images, and cytotoxicity genes expression, like IFNG, PRF1, GZMB, will be run on real-time qPCR. CD3 will be used as housekeeping gene.
I know the PHA will activate T cells and stimulate them proliferating via CD2 and calcium influx pathway. And I do observe obvious cytotoxicity in the preliminary experiments. Could anybody give me some advices to improve this method?  . 
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I think that 3000 cells are not enough to isolate RNA to run qPCR.
Plus, cytotoxicity of T cells is not very strong and 6 hour of co-culture maybe is not long engough.
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Dear altruist,
I am working on Bioinformatics and Computational Biology. I am using Cytoscape tool for network visualization or analysis. Now I am working on lung cancer-related genes. In this work, I want to add some clustering methods to analyze for a better outcome. Please suggest me which one is better than any other plugin?
I already use the clusterOne plugin but want something more such like as MCL, k-medoid clustering.
Thanks for advance.
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Hi
you can find here a list of plugins that can be used for clustering on Cytoscape.
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for example, I have used POPE cell membrane in my research work (molecular dynamics), but according to the paper that I attached below, this membrane is a kind of normal membrane.
if there is somebody who knows how to modify this membrane as a cancerous membrane, please recommend me.
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I would suggest you to look for a litterature about the lipid composition of cancerous cell membranes. POPE is just a model-membrane which does not need to be a representative for a cancerous cell membrane. Moreover, from the behaviour of compounds with the membrane composed only of POPE lipids you can not draw conclusions of the behaviour of cancerous cell membrane. To do more accurate research you shall also keep in mind what kind of cancer you are dealing with? In what tissue? Depending on tissues you will find the lipid composition which you need.
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I tested 25 ug/ml and 40ug/ml of Fluorescently labeled transferrin in melanome cells during different times of observation, but no fluorescent signal was detected inside the cells.
Is there anyone here who has had the same problem?
Thank you.
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I'm coming a bit late but I also have trouble with transferrin, I can't detect any signal after using the concentration given by the literature. Did you try to use it at a higher concentration on the HDF cells? Do you remember if it worked ?
I do thank you
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The plant is already known as anti-cancerous, but the biochemical study has not been done. However the active compound, from the plant has been researched already to find anti-cancerous activity in various in-silico studies, however no wet-lab is done. How should the road map of the studies can be carried out ?
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Test the raw extract on cancer cell lines in vitro and chemically induced cancer in vivo, if the extract. If it showed anticancer effect, try to separate and purificate the effective ingredients and test them separately. Determine the LD50 of the effective compound in two animal species. Perform preclinical study. Then clinical study
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I am working on Epithelial to Mesenchymal Transition (EMT) in A549 cell line by treating positive inducers such as EGF to up-regulate the mesenchymal markers like vimentin. I'm doing western blot to detect the changes of vimentin expression level over treatment time of the inducers. I could detect the protein band of vimentin at the position of its proper size, but the change of protein expression level flutuate too much.
For example, I expected increase of the vimentin expression level by the treatment of EGF at least in early timepoints right after the treatment, but it wasn't. Neither at longer imepoints. Sometimes the level of vimentin was completely disappear, sometimes increased, or just remained in the same level. I always confirmed the expression level of beta-actin or GAPDH as a loading control, and it was consistent.
I tried same kind of expreriments using mesenchymal cell line like MDA-MB-231, and in this case, the degree of the fluctuation was decreased but it doesn't completely disappear.
The passage of the cell lines are all around 10, which is I think low enough, and I grow my cells in complete medium RPMI 1640 (10% FBS, 1% Penicillin/Streptomycin) and I treated the EMT inducers after 16 hours serum starvation.
I doubt that the A549 cells that I prepared for the experiments were not in good condition, so this can be the reason of the fluctuation. Do you think that the health of the cell can affect to the level of vimentin much? If so, how can I troubleshoot my problem? Any suggestion or advices can help me a lot. Thanks in advance.
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The problem is with gel concentration and high molecular weight of Vimentin. Use 8% gel and try to load 40ug protein instead 20ug. I hope you will get it. I have worked with EMT and got Vimentin blot in my master thesis. A549 I think it is NSLC cell line. Our previous lab has done , no problem with cells as well. Good luck.
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I would like to know the viscosity of Matrigel (Corning) as function of temperature. I already know that it solidifies at 37° and its liquid at 4°, but i would like to know what's the transition form, and what happens with different Matrigel concentrations.
Thanks in advance,
Simon
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I had put some of the Matrigel I was culturing my HUVECs on on a rheometer a while back, and it had produced a pretty flat G'/G'' from 20ºC to ~40ºC. I don't exactly remember the details, unfortunately.
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Mack et al studied subtypes of three ependymoma(same histopathology) brain tumors and found that one subtype carries an intrachromosomal translocation that creates a new tumor-driving gene, another lacks tumor-driving mutations but has aberrant epigenetic modifications, and a third shows neither gene mutations nor epigenetic aberrations. There were three genotype but one cancer phenotype. Similarly Martincorena and colleagues found thousands of mutations in cancer-relevant genes, including cancer-driver genes, in normal eyelid epidermis .(multiple cancer genotypes but no cancer phenotype).
In disparate classes of biological systems, there are more genotypes than phenotypes. Where sufficient information exists to enumerate these phenotypes, there are exponentially more genotypes than phenotypes, as a function of the number of system parts. This means that any one phenotype typically has many genotypes that form it.
In a brief, cancer is the decision of the cell to choose the innovative/adaptive phenotype and understanding the genotype does not mean understanding cancer.
References
1. Mack, S. C., Witt, H., Piro, R. M., Gu, L., Zuyderduyn, S., Stütz, A. M., et al. (2014). Epigenomic alterations define lethal CIMP-positive ependymomas of infancy. Nature 506, 445–450.
2. Martincorena, I., Roshan, A., Gerstung, M., Ellis, P., Van Loo, P., McLaren, S., et al. (2015). High burden and pervasive positive selection of somatic mutations in normal human skin. Science 348, 880–886.
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Answer of the question
Cancer is name of the a kind phenotype not genotype
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I am doing a pilot study to assess the cytotoxic abilities of different T lymphocyte populations when cocultured with B16 Ova In Vitro. I have seen different articles labeling their effector/cytotoxic T Cells with a target dye to gate them out and look at Annexin and PI positive cells. But I do not have any target dye to label my cells. Instead can harvest my cocultured tumor + T cells and first do a CD4 CD8 surface staining with antibodies and then follow Annexin PI staining protocol? I understand the 2 stainings are different but will that affect my apoptosis results in any way? Has anyone done this before and has had luck with it? Please help me to understand better. Thanks
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Hi Sarah Ratkovich-Gonzalez , I remember that you shoul perform the surface staining first and then stain with AV (and 7AAD for example). That is because for AV staining you need to you a "special" binding buffer.
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During routine microscopic observation of Cancer Associated Fibroblasts (CAFs), my colleague Pedro Barcellos and I came across these odd geometric "dark" structures inside the cells. We haven't been able to determine what this would be. This was not evident in all cells, but in a fair share of them. And also present in different biological replicates.
We would love to know what these structures represent or to have any hints about what they might be.
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Murilo,
Very interesting observation in your fibroblasts! For years I thought the accumulations of F-actin might be due to some defect in cytoskeletal remodeling in the exostosis chondrocytes. Perhaps there is a block in depolymerizing into actin monomers, which might be associated with or caused by the excessive bundling by alpha-actinin and presence of alpha-actin which normally should be in muscle. I really hope you solve the mystery and look forward to your findings in fibroblasts.
Please also click the "recommend" button if the papers or comments are helpful. Thanks, and best of luck.
Mark Bernard
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In collaboration with Frontiers in Oncology (latest Clarivate Analytics' impact factor: 4.416), "Cancer Metabolism" section, and together with Professors Athanassios Vassilopoulos and Lucia Altucci, we are bringing together a selected group of international experts to contribute to an open-access article collection titled "Sirtuinome Rewiring to Hijack Cancer Cell Behavior and Hamper Resistance to Anticancer Intervention". Frontiers provides a free and immediate Gold Open Access-based online access to the scholarly literature for anyone in the world to read, distribute and reuse. In this regard, we are pleased to inform that a 25% discount will be granted for accepted manuscripts. For more detailed informations, please go to https://www.frontiersin.org/research-topics/10447, and remember that submission deadline is November 2nd, 2019. ------------------------------------------------------------------------------------------------------------- Call for Papers Extensive reprogramming of energy metabolism and detoxification processes are increasingly seen as critical factors involved in metastatic progression and in development of chemo- and radio-resistance. Mammal sirtuins (SIRT1-7) are a family of conserved NAD+-dependent protein deac(et)ylases and/or mono-[ADP-ribosyl]transferases with varied cellular distribution. Their role as crucial regulators in energy metabolism and adaptation to cellular stress is currently under extensive investigation worldwide, not only in physiological processes (e.g. in aging) but also in the pathogenesis of cardiovascular and neurodegenerative diseases, diabetes and cancer. In particular, sirtuin-dependent signaling is suspected to play a dual role in cell biology, on one hand protecting DNA from genomic instability and limiting the replicative potential, on the other hand inhibiting senescence and promoting survival and growth advantage. Interestingly, SIRT3-5 localize to mitochondria and regulate targets involved in a diverse array of biomolecular pathways, many of which govern energy metabolism and apoptotic death. Such characteristics confer a great importance to sirtuins, in terms of preventive medicine and therapeutic potential. Unfortunately, despite the rapidly increasing interest in the field, results are still insufficient to draw definitive conclusions. More importantly, the question as to whether sirtuins can be considered as tumor suppressors or oncogenic proteins remains unanswered. In this Research Topic we welcome Original Research and Review articles focused on clarifying the mechanisms underlying sirtuin-driven responses to endogenous and exogenous stressors in tumor and malignant cells, in terms of metabolic rewiring, antioxidant protection and cell cycle control. In addition, researchers are also invited to provide data and opinion regarding strategies aimed at controlling the expression and activity of sirtuin-responsive cellular systems, with particular attention to pharmacological and nutraceutical approaches. Finally, studies focused on sirtuin-dependent pathways leading to malignant progression and/or development of chemo- and radio-resistance in cancer cells are also welcome. Potential topics include but are not strictly limited to the following: 1) In vitro or in vivo studies focused on sirtuin-dependent regulation of redox-based responses to stressors in tumor and malignant cells, with particular attention to metabolic reprogramming, DNA damage repairing capacity, antioxidative potential, and proliferation; 2) In vitro or in vivo researches on the mechanisms underlying the action of synthetic and natural compounds on the expression and activity of sirtuins and downstream cellular pathways in tumor and malignant cells; 3) In vitro or in vivo studies investigating the role of sirtuin-dependent intracellular and extracellular signaling associated with cancer progression and/or development of resistance towards chemo- and/or radio-therapies. Stefano Falone, Ph.D. (Department of Clinical Medicine, Public Health, Life Sciences and Environment Sciences, University of L'Aquila, L’Aquila, Italy) Athanassios Vassilopoulos, Ph.D. (Northwestern University, Chicago, IL, USA) Lucia Altucci, Ph.D. (University "Luigi Vanvitelli", Naples, Italy) Topic Editors, Cancer Metabolism Section, Frontiers in Oncology
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So, if there is a rescue, it may be challenging to characterize.
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It is confusing concept in the literature that some papers use the term "HER2 positive" and some "HER2 overexpression". You may refer to the definition in the specific paper, but sometimes they even dont mention what they mean with "HER2 positive" cell.
Any clarification would highly appreciated.
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It depends on the cell line. "Overexpression" can be defined as a state in which the expression is higher (e. g. after a treatment) than in the"normal" state - this needs to be compared.
Hope this helps
WD
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What are the best and (affordable) techniques for the Mesenchymal Stem Cell isolation from mice or rat cells especially using on cancer studies?
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In our publication we used wharton jelly fragments as avery rich source of MSC
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Cancer cells uptake of NPs time period?
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Hi, I agree with the Sanjib Saha answer for this question
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I'm a Research associate at 57357 hospital , Egypt , I would like to ask about two cell lines : RD , RH30 ( Rahbdomyosarcoma ) we couldn't find them in Egypt and we are committed to certain budget , deadlines and masters thesis based on this project , anyone know a trusted source  or a lab in any country usually uses them to give as aliquot ? Thank you in advance !
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Has any serum protein been associated with the induction of EMT/MET of solid tumor cell?
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Joe Graymer
that is an interesting perspective
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what is the best protocol to culture lymph node from fresh tissue sample ? best media , best serum concentration , any other additives to be added ?
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Amr Mahmoud RPMI is generally used for culturing lymphocytes, I've grown primary T cells and B cell lines in RPMI and they are fine. I have not used these media for generating cell lines so I can't comment on which is more suitable for your procedure. AIM-V might be a good shot for lymphocytes if you wish to avoid bovine serum supplementation. Good luck.
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I found this interesting device (https://darwin-microfluidics.com/collections/microfluidic-organ-on-a-chip/products/ddi-chip-distance-dependent-interaction-chip-pack-of-3). Have you any experience in the use of microfluidics to study cell cross-talking and signaling?
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