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Colon Cancer - Science topic

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Questions related to Colon Cancer
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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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Hi guys,
Do you know for how many passages you can keep the cells CCD 841 CoN in culture?
Best
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Dear Good people,
Is it applicable to perform in vitro assays on a Particular cancer cell line that differs from a normal cell line?
for example to work on colon cancer cell line and have normal liver cell line as a control!
Thanks In advance
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Let me explain to you with an example.
If you are going to work on invitro cytotoxicity assay using colon cancer cell line to test the cytotoxic effect of the candidate drug, you will use a drug that is already known to cause cytotoxic effect on colon cancer cell line, as a positive control.
Your negative control for this invitro assay will be colon cancer cells left untreated with the candidate drug.
You will not use a normal cell line as a control in this assay.
Usually, a normal cell line is used to test the cytotoxic effect of the candidate drug on normal cells. This is done when one is interested in calculating the selective index (SI) of the candidate drug. SI will indicate how selective is the candidate drug in killing the cancer cells.
SI= CC50/IC50
Where,
CC50 is the concentration of the candidate drug required to reduce cell viability by 50% using normal cell line.
IC50 indicates how much candidate drug is needed to inhibit a biological process by half using cancer cell line.
The higher the SI ratio, the more effective and safer is the candidate drug.
Hope this will help answer your question!
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I want to compare human colon cancer gene MLH1 with mutS sequence and other related genes.
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Also check please the following useful link:
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Hi all,
I am thawing some HCT-8 cells from ATCC and it calls for RPMI with horse serum. Does anyone know the reasoning for this and if they will tolerate FBS?
Thanks,
Claire
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HCT-8 colon cancer cells are human adenocarcinoma cells from the ileocecal region which may give a reflection of gut absorption. The cells produce a polarized differentiated monolayer consisting of enterocyte-like columnar cells that are interconnected by mature tight junctions and desmosomes and exposed microvilla.
For normal cell culturing you could use 10% FBS. But if you need to switch HCT-8 cells to differentiation state, you may accomplish this by supplementing the medium with horse serum instead of FBS. You could either use 10% horse serum or 5% horse serum along with 5%FBS which will help to slow down cell proliferation considerably.
When you maintain cells in culture, FBS is generally used because FBS contains factors for proliferation that will help cells to proliferate. Whenever cells have to be switched to differentiated state, they have to be withdrawn from the proliferative state.
Best.
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Theoretically, Stage III includes IIIA, IIIB, IIIC. So, I wonder why is there a fourth group (Stage III) in the Subtype profile workflow at Gent2 database.
My other question does anyone know a database where mRNA and protein datas from blood samples are available from cancer and normal patients?
Thanks,
Anita Kurilla
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There is no yield of specific tyrosine kinase inhibitory activities (VEGFR, c-Met etc.) of my molecules. (General tyrosine kinase effect is known). Is it a correct approach to correlate the anticancer activities on kidney, breast and colon cancer (the order of molecules according to activity is different in each cancer line) with some of the receptor tyrosine kinases (PDGFR, EGFR) of my choice? For example, can I correlate binding affinity in PDGFR with anticancer activity in colon cancer based on compatibility? According to which receptor affinity is compatible with activity in which cancer line.
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TNF, CRP must be taken into account for anticancer activity
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This is the human/Control tissue for IHC staining. I have another one for treatment group. what is the solution for this type of tear of tissue in IHC.
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Possible reasons/solutions:
1) Bad FFPE sample quality, likely bad fixation.
2) Bad sectioning - use fresh blade, soak trimmed block surface before sectioning on wet (dH2O) tissue for 5-10 min (preferred method in our lab), some people cool blocks before sectioning.
3) Bad slides. Use proper ones. I routinely use Superfrost Plus from Fisher, cat 12-550-15
4) Before deparaffinization, incubate slides at 50 or 60 Celsius for 30 minutes. It may help when you have harsh antigen retrieval conditions.
5) To aggressive antigen retrieval (AR): boiling time, temperature, microwave exposure time, concentration of enzymes.
If your slides before deparaffinization look intact under a microscope – this is a perfect. Just incubate slides at 50-60*C for 30-60 min and be gentle with AR
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I want to cote a tablet for colon targeting. Which is the best agents for such task? because pH varies from stomach to colon, so which type of agent is best for such operation?
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drug delivery system is novel drug delivery systems which has an upper hand owing to its ability of prolonged retaining ability in the stomach and thereby increase gastric residence time of drugs and also improves bioavailability of drugs
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To achieve the goal of personalized medicine it is necessary not only to have agents with defined molecular specificities, but to have minimally invasive biomarker and imaging tests that will identify which patients have the target in their tumor and the patient’s pharmacogenotype. A related objective is to have a way to measure the effect of a drug on its molecular target in the tumor to be able to answer important questions, such as how much drug is required to inhibit the target in a tumor and is there is a benefit to giving more drug or will this only increase toxicity.
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The Oncotype DX portfolio of breast, colon, and prostate cancer tests applies advanced genomic science to reveal the unique biology of a tumor in order to optimize cancer treatment decisions. In breast cancer, the Oncotype DX Breast Recurrence Score test is the only test that has been shown to predict the likelihood of chemotherapy benefit as well as recurrence in invasive breast cancer. Additionally, the Oncotype DX Breast DCIS Score test predicts the likelihood of recurrence in a pre-invasive form of breast cancer called DCIS. In prostate cancer, the Oncotype DX Genomic Prostate Score® test predicts disease aggressiveness and further clarifies the current and future risk of cancer prior to treatment intervention.
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Dear colleagues,
I am studying mouse NK cells infiltrating MC38 colon cancer or B16 melanoma inoculated into the syngeneic C57B/6 mice. To this end, I need to identify the NK cells that are naive, inhibited, activated, post-activation, memory, exhausted, etc. What are the best (surface) markers for these subsets? Any advice highly appreciated.
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We didn't use this on NK, only use this on T cell for the exhausted marker. If you want to identify the activated NK and exhausted NK , you may try to evaluate some cytokines or relevant downstream proteins.
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I´m trying to establish a coculture with colon cancer cells, which are adherent cells, and PBMC, non adherent cells. So I would like to get a direct contact coculture, I mean, without barrier between the cell populations, but I have seen that the cancer cells get detached when they are in contact with the PBMCs. So I would like to know if anyone has made this coculture and what could be a suitable method to get it.
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I am trying to do a similar experiment. Waiting for any response. My plan to seed different densities of adherent cancer cells for at least 24 hrs before adding PBMC cells. Then, I will see the effect and at which confluences the effect will be starting.
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Am working on a DMH induced colon cancer in mice and i would like to know, could polyps or ACFs be counted through visual macroscopic examination without methylene blue staining ?
Thank you.
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Hichem Moulahoum Thank you the detailed answer.
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I want to try to induce colon cancer in mice, and then evaluate potency of anticancer drug in these mice. It means I will have to evaluate size of tumor following injection of antitumor drug in several days .
Is there anyone who induce this tumor in mice? Does it have especial protocol?
Thanks
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Injecting the dimethylhydrazine and after two months will develop the cancer
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I used EGF to make EGFR activation on HCT116 colon cancer cells in the cell incubator for 10 mins, then BS3 crosslinker (Thermo Fisher) was employed to EGFR dimerization. I tried to different ways for BS3 incubation: at room temperature for 20mins and 40mins or on the icebox (I'm worried that EGFR internalizes into cells so I put it on the ice to reduce the internalization), 250mM glycine for quench. Finally collecting the samples by 4X SDS buffer (without 2-mercaptoethanol) and ready for immunoblot. I can see the EGFR monomer expression (170KDa) on the membrane but I can't see dimer. I used two different anti-EGFR antibodies from Cell signalling and PeproTech and the EGFR dimer still not appear on my membrane.
Can anyone whoever did the Western-blot for EGFR dimer tell me what should I revise for my protocol or anything else I can do for Dimer detection?
Thanks a lot.
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Yes you can increase it, but it may be due to too little EGF sensitization. I would test more EGF first, or longer incubation time with EGF before I alter BS3 for it is more likely that there aren't enough dimers forming than BS3 failing to crosslink them.
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I am confused as to which ELISA kit type to use for a colon cancer work, Sandwich or Competitive and why?
Thank you
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For your purpose, the sandwich is the best.
competitive is often for small molecules test.
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How to get access to LYON19 dataset? histopathological image dataset of Lymphocytes provided by LYON19 challenge is not accessible from its challenge site https://lyon19.grand-challenge.org/Background/ .
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Train dataset not available in this chalange..
" No training set is provided, participants should use their own data to develop a method. "
Test data can be downloaded from the  Zenodo platform
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Our approach to R Hemicolectomy is starting from mid point of Transverse colon Gastro colic ligament using an articulating Enseal,Entire t colon can be mobilized with out frequent change of instruments,much faster,equally effective mobilisation and no added problem to duodenum and ureter.Please see the Link for the technique.
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I don't use it but it may be of some interest in the case of D3 lymph node dissection as the retroperitoneal mobilisation of the right colon allows easier dissection along the superior mesenteric vein axis and skeletonisation of the right colic branches.
I agree that the tenet of "no touch technique" prioritizes vascular ligation before mobilisation of the colon. Nevertheless, we have to accept that no data exist showing a survival benefit for laparoscopic medial to lateral, lateral to medial or D3 lymph node dissection in right colon cancers.
Prasanna Kumar Reddy do you continue using the craniocaudal dissection or have you shifted to another technique?
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"In chronic inflammation, IL-6 has a detrimental role that favours mononuclear cell accumulation at the site of injury, through continuous MCP-1 secretion, angioproliferation and anti-apoptotic functions on T cells [30]. This may increase serum levels of IL-6 and provide the basis for the amplification step of chronic inflammatory proliferation."
Interleukin-6 is released by monocytes and macrophages in response to other inflammatory cytokines which include interleukin-11, and tumor necrosis factor (TNF)-beta.
"Chronic inflammation can result from the following:
  1. Failure of eliminating the agent causing an acute inflammation such as infectious organisms including Mycobacterium tuberculosis, protozoa, fungi, and other parasites that can resist host defenses and remain in the tissue for an extended period.
  2. Exposure to a low level of a particular irritant or foreign materials that cannot be eliminated by enzymatic breakdown or phagocytosis in the body including substances or industrial chemical that can be inhaled over a long period, for example, silica dust.
  3. An autoimmune disorder in which the immune system is sensitized to the normal component of the body and attacks healthy tissue giving rise to diseases such as rheumatoid arthritis, systemic lupus erythematosus (SLE).
  4. Recurrent episodes of acute inflammation. However, in some cases, chronic inflammation is an independent response and not a sequel to acute inflammation for example diseases such as tuberculosis and rheumatoid arthritis.
  5. Inflammatory and biochemical inducers are causing oxidative stress and mitochondrial dysfunction such as increased production of free radical molecules, advanced glycation end products (AGEs), uric acid (urate) crystals, oxidized lipoproteins, homocysteine, and others."
Cytokine Panel (ARUP):
Interleukin 2 Receptor (CD25) Soluble
Interleukin 12
Interferon gamma
Interleukin 4
Interleukin 5
Interleukin 10
Interleukin 13
Interleukin 1 beta
Interleukin 6
Interleukin 8
Tumor Necrosis Factor - alpha
Interleukin 2
Interleukin 17
Question:
If CRC patients had a hair sample analyzed for fungal cultures, what fungi would be found?
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Bogdan Socea Thanks for sharing - I agree!
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I am writing a paper on "colon cancer and nutrients". While searching for literature a point hit my mind: can bacterial infection be one cause of colon cancer?.
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I am currently using CT26.wt cells colon cancer injected s.c in the flank of Balb/c mice (synergic model) to study effect of a treatment on the colon cancer in mice. I used 0.2x10^6 cells to establish tumor. According to my study design, the end point is when the tumor volume reaches (1000mm^3).
Comparing to the treated group, the control group reaches the end point faster than the treated ones which make impossible to get the tumor picture simultaneously to prove the effectiveness of the drug. Is there any way to prove (except the tumor volume chart and fluorescence image)? Thank you in advance.
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If you are focusing on the effect of a novel agent, maybe you can do the evaluation by calculating T/C% of each time point and finally make a significance analysis.
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I am growing spheroids from HT29 colon cancer cells, and I need some sort of control cells to be compared with the HT29. Now, I should grow the control cell into spheroids and then use these new spheroids as control for metabolic rate, for instance. I found several normal cell lines, both fibroblasts and epithelial cells, but I do not know if these cell lines (CCD-33Co, FHC, CCD 841 CoN, CCD-112 CoN, CCD-18Co):
- 1: form spheroids;
- 2: are good as control cells to compare metabolic rate with a cancer cell line as HT29;
- 3: what is best between fibroblasts and epithelial cells for my application;
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Hi Pilar,
it's possible to grow spheroid from primary cells but you need to force their aggregation in spheres by using V-shaped ULA wells. We have tested in our lab.
Feel free to contact me in private message to go ahead with the discussion
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Dear all, we are evaluating the cytotoxic effect of plant extracts on HT-29 and HCT116 colon cancer cells. We have determined the IC50 values for each cell line, however, we want also to test those concentrations in the normal colon cells (we are going to use the CCD-33Co cell line) to discard toxic effects on these cells. Which should be the concentrations that we must test in these cells? Some people have suggested us to use up to 3 times the IC50 values but we haven't found documents justifying this. Could you please provide documents or papers indicating suggested concentrations? Thanks in advance for your help.
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Es eso a lo que me refiero con lo de adoptar criterios toxicológicos. Los alcances de un abordaje in vitro no pueden sustentar la inferencia de inocuidad, así haya una tolerancia 3x superior a la presentada por sus contrapartes de cáncer (atributo que no comparte ningún agente o candidato a quimioterapéutico). Por la sencilla razón de que la toxicidad de un fármaco (o candidato a) debe ser estudiada a escala organísmica y nunca a nivel células aisladas, por su pobre representatividad de una hipotética administración a individuos.
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I'm doing western blot to detect LRRC8A protein which is extracted from colon cancer cell HCT116, unfortunately, I could not find a good result. Does anyone have an idea or advice?
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Hi Shada,
Without seeing your results, I can just assume that your antibody is not a monoclonal antibody. Find a monoclonal antibody for your protein. When it is not the case, please share your result. If you can add one image from your western blot results, one could evaluate your problem better.
I hope this answer helps you.
Cigdem
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Green chemistry ,and green organic food is proved healthy,but many peoples dont recognize and take these topic for serious .Most disease.irrited colon and cancer come from inorganic and cholesterol diets,so all research goes to organic food and plant and vegetables But why many peoples dont take this as serious?.
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Following
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Phytonutrients properties of functional foods against colon cancer-inducing agents, such as azoxymethane, in animal models
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I'm trying to compare caloric intake of patients with colon cancer with the calculated ideal caloric intake. What statistical test should I use? assuming data are normally distributed should I use paired t-test?
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I found this answer very relevant to the question:
Statistical methods for numerical variable
there are two groups: 1. comparison group(T-test and ANOVA)
2. correlation and regression
t-test(paired and un paired )
UNPAIRED: the data are independent that is one nominal level variable (two different groups/categorical)with two group as independent variable that are mutually exclusive .eg. chomper mean birth weight between males and females
by independent of data we mean that the data value of each study subject rises independently uninfluenced by and uncorrelated with the data values of the other subjects. the distribution of the dependent variable /numerical variable normal(normally distributed). the test also assume that the variance (of the dependent variable) in the two groups are equal. the assumption is called the requirement of homogeneity of variance
PAIRED: data s are side to be paired , if the study subjects from one population can be matched or paired with particular subjects in the second population. paired data rise naturally from studies of twins and paired objects such as eyes or ears of the same individual.
advantage of paired data is that smaller size are needed b/c of the similarity , hence decries variability with in paired .paired data also arises in studies where observation are taken before and after the intervention on the same study subject.
paired data can be analysis the d/c b/n each member of the pair and tests to determine if the difference are significantly d/t from zero/ the data are normally distributed. eg. com paired mean blood pressure of diabetic patients before and after some intervention/treatment . one group matched twice rather than two independent group.
but your out come variable is categorical (case and control) not possible to use independent 2 samples or paired samples T test (it is wrong method of analysis)
The dependent /outcome/ response variable is categorical the appropriate method of analysis.
your out come variable is categorical , the baste method of analysis of your study or data is:
1. chi squared test of Independence( to check the association between categorical dependent variable and continuous or categorical independent variable)
2.matched paired test(cross-sectional , case control the data s are independent sample / before and after the study cross over or matched case control studies) each cell represent the number of pair in which both member of pair experienced the same value with the two equal to the number of pair
3. in paired sample we use MCNemar's test is the proper method of analysis to test the hypothesis
4. conditional logistic regression ( bi variate and multivariable analysis of logistic regression ) to check or to identify factors that affect the dependent variable(case and control) through controlled factors(independent variable) and confounding.the crud and a dusted OR together with the corresponding Confidence interval computed. A good fit as measured by Housmer lemeshow's test.
How to Select the Appropriate statistical Analise Selection criteria for statistical tests
First, you have to define the level of measurement of each variable to be included in the analysis.
Second, to select the correct statistical analysis, you have to clarify what you want to find out.
Third, sample size calculation or power analysis is directly related to the statistical test that is chosen.
The selection of a statistical test is based on the purpose of the test, the experimental design, and the type of variable (generally, measurement or rank).
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I am doing research on colon cancer Want to compare two different treatments of survival in a retrospective data. I want to do propensity score matching to adjust for the bias. I am using inverse propensity score weight method. I calculated inverse propensity score weight from propensity score. I was able to get odds ratio by using this weighted variable in SPSS. But when i am trying to calculate KM curve I am getting the following error code "No statistics are computed because nonpositive or fractional case weights were found". Any suggestions of how to proceed? 
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Would you like to provide R code for IPTW?
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Looking to understand effects of ROS at different stages of colon cancer.
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Thank you!
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What we know:
So, what percentage of physicians actually order KRAS genetic testing for their colon cancer patients? To be determined.
Is it cost effective? Turns out it's average.
"Results
Screening for both KRAS and BRAF mutations compared with the base strategy (of no anti-EGFR therapy) increases expected overall survival by 0.034 years at a cost of $22 033, yielding an incremental cost-effectiveness ratio of approximately $650 000 per additional year of life. Compared with anti-EGFR therapy without screening, adding KRAS testing saves approximately $7500 per patient; adding BRAF testing saves another $1023, with little reduction in expected survival.
Conclusions
Screening for KRAS and BFAF mutation improves the cost-effectiveness of anti-EGFR therapy, but the incremental cost effectiveness ratio remains above the generally accepted threshold for acceptable cost effectiveness ratio of $100 000/quality adjusted life year."
To Consider:
(1) What drugs might be more effective against colon cancer cells bearing the KRAS mutation?
(2) What drugs might be more cost effective against colon cancer cells bearing the KRAS mutation?
(3) Relating to cost effective treatment, how often do we prescribe drugs or assign treatment plans that are expensive $$$, decrease the length of the patient's life, and decrease the patient's quality of life? How can this be prevented? (E.g., recommending surgery procedures for aged colon cancer patients). How do we incentivize treatment that is most cost effective?
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How would I go about investigating state of the intestinal epithelium of someone with IBD and colon cancer?what would I observe f I use immunostaining
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With inmunochemical studies about the lymphocytes subtipes and several interleukines perhaps
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In this article fig. 1. The Author has made a decision tree with 24 nodes. Each node is specified for a specific cancer tissue of origin and the couple of MicroRNA which can identify these cancer tissues of origin. My question is, if I isolate miRNAs at the node14(hsa-miR-21, let-7e), node21(hsa-miR-205, 152), node24 (hsa-miR182, 34a, 148), node10(hsa-miR-194, 382, 210), will it be enough to identify cancerous tissue originated from the lung.
Why am I asking this question, because, I want to identify cancerous tissue, which has migrated to different region but originated in the lung. So if I take miRNAs from those specified nodes, will it be enough to identify lung cancer tissue, which has migrated to different region but originated in the lung.
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microRNAs have to provide potential signature model for various cancers and other diseases. While miRNA in terms of sequencing is difficult , though being done provides huge number of miRNA in a specific cancer as you have exemplified some in your discussion. Few important learning points:
1- Specific sets of miRNAs usually express or otherwise together and they are termed miRNA families.
2-Some miRNA have the potential to act as pan cancer biomarkers but still no specificity is provided.
3-Some biomarkers are like positive or negative acute phase reactant and may rise or fall with non-cancerous disease.
4-There is some but as i experienced little correlation between blood and tissue based miRNAs
5- The science of miRNAs is still emerging and a lot more has to be learnt i guess before they are available, if available for clinical use.
6-Also need to have complete data about pre, pri and miRNA and he cleaving proteins like DROSHA, DICER and factors incorporated in RISC complex.
The whole picture is yet to appear and but hope is there that someday they may be appearing as both for diagnostic use and therapeutic targets like Riversin (spelling ?) for treating hepatitis C.
So potential is there but more research is needed to quantify and deal associated aspects of miRNA
Sorry, that I could not help you straight as i interpret the knowledge about this subject is still evolving.
Kind regards
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Hi all,
Did anyone worked on developing spheroids of 0.8 - 1.0 mm size from colon cancer lines.
Please help me in some techniques to handle spheroids
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Thank you for your answer.
May i know the reason why you are advising to go with reduced serum levels.
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colon cancer proliferation experiment I use control groups for this (I do not add chemicals to the control groups) but in my control groups I die as groups that I add chemicals. I need to set a dose, but I can't determine the dose because they're dead. Do you have any suggestions for me?
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CCl-233 colon cancer
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Can anyone help me with my research (mRNA expression of Caspase-3 on colorectal cancer) please?
I need help to understand my results.
Thanx in advance
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Thank you
normal group has low level of caspase-3, does it right?
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It is a known fact now that interactions between bacterial (normal flora) and colon cancer cells influence the transcription profile of host cells or enterocytes.
So, how can I avoid or minimize or cleanse the intestinal bacterial load of human colon biopsies stored in RNALater at the time of surgery, before proceeding to RNA extraction from them to avoid at maximum all the bacterial RNA burden during my extraction for the human RNA. RNALater is a well-known bacteriostatic but not the bactericidal. Is there any sound protocol exist to tackle this issue or this bacterial RNA load is insignificant for various gene expression analyses over such colon samples?
Though my primers are quite specific, but still I have concerns. Need some deep insights or explanation about this scenario.
Thanx...
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I suppose its practically impossible to get rid of 100% of bacterial RNA by any kind of cleaning. If the primes are specific the bacterial RNA should not interfere with your analysis.
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I tried to induce colon cancer in male mice, following a paper that used Cyclophosphamide in a dose of 40 mg/kg (i.p.), 5 days of injection followed by 30 days of incubation.
But that did not work. I tried to increase the dose, but mice died.
Is there anyone who used CYP before in such thing? or is there any efficient alternatives for induction of colon cancer in mice?
Thanks
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I work with CT26 model in NRC, in case you are interested, please let me know.
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I found the MC38 cell line in our liquid nitrogen storage system. The origin is unknown for the moment and the cells have been stored away for quite some time.
I would like to perform some preliminary experiments on these cells before purchasing them.
Is there any way I can screen these cells to ensure they are actually MC38 colon carcinoma cells?
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Lukman Afolabi We were unable to trace the exact origin of the cells, so I only kept them in culture for several weeks. During this period, the cells did not expand very quickly, however I found the rate acceptable. If I'm not mistaking, I used a split ratio of 1:5 from Monday to Wednesday.
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Gain-of-functional mutant p53 exhibits a longer half-life period as compared with wild-type p53, which is why the significant accumulation of the mutant p53 in cancer cells is recognized in many kinds of malignant neoplasms.
As compared with CEA and CA19-9, anti-p53 antibody (IgG subtype) can be detected in the very early stage of esophageal, colon, breast, prostate carcinomas etc.
Given that pseudo-positive ratio of anti-p53 antibody is less than 5% in the healthy population, the detection of anti-mutant p53 IgG antibody holds much promising.
I would like to know your opinion on this useful marker.
Thank you in advance!
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I would like to add that anti-p53 antibodies have been reported in other human diseases (other than cancers) such as auto-inflammatory and autoimmune diseases. Therefore, reducing its clinical usefulness.
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Capecitabine solubility in water
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About such substances they say that they are diphilic or amphiphilic. Diphilicity is determined by the partition coefficient between octyl alcohol and water. There are tables of coefficients of distribution of a large number of substances and approximate calculations of these quantities for the chemical structure of matter.
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Good day all, I am about starting an experiment, in which I have to induce colon cancer.
(1). Please can anyone tell me the best model to use among these; Methylazoxymethanol (MAM), 1,2-dimethylhydrazine (DMH), and azoxymethane (AOM). And also, how long will each of these agent take to induce colon cancer in rats.
(2). Can 1,2-dimethylhydrazine and azoxymethane be combined?
(3). What is the best age to induce colon cancer in rats for both sexes?
I would be grateful if anyone can help.
Thanks in advance
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Thanks for your informative answer
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I want a choose a/some antibiotic to disturb mouse gut microbiota to explorethe effect of gut microbiota disorder in colon cancer liver metastasis, I have looked up some literatures and many antibiotics were choosed,and I have no idea which one to choose. Which antibiotic has more significant of pathophysiology. I want the antibiotic that make effect is similar to pathophysiology and will occur in fact ranther than a environment I give.Help~
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I suggest enrofloxacin 5 mg/kg SC.
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Hi everyone,
I'm working with immune response in colon cancer patients. Now I just wonder about that can I measure the NK cell activity? 
Thank you in advance
Da
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See this link of article is useful
Good luck
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Hello,
I have been performing live-dead staining in my colon cancer primary cells in order to access viability after different treatments.
I found that PI shows up all over the cells, either they are live or dead. PI should only stain the nucleus of dead cells, so this is intriguing. When taking pictures using the GFP channel (for FDA staining) and PI channel (for PI staining), the raw pictures are exactly the same.
This is happening to other colleges of mine that use the same protocol in different cells. In live imaging, the parameters can be adjusted in order to obtain a brighter red and have an idea of how many dead cells, but this is not an accurate method and I cannot used to quantify anything.
Before buying another expensive dye, I would like to know if someone has some ideas about what is happening...
Thank you in advance.
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Hi Ana,
Have you tried Acridine-orange and ethidium bromide dual staining method? It really works well, at least it worked for me while staining cultured cells.Both of these dyes are rather cheaper than the other fluorescent dyes/kits available for live-dead imaging. But, I am not sure whether or not they would work in primary cells. You can give it a go!
Have you tried trypan blue, anyway? I am not sure though about this.
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I am basically a chemist and I am culturing APC deleted colon cancer organoids. I want to know how to isolate DNA from them. Can anyone suggest?
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It is better to use Qiagen, but you can use any kits to extract DNA from the tissue. For example, Tiangen, Roche, Thermo Fisher and other.
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I need a dataset (images of a colon biopsy sample) to diagnose and grade colon cancer. I did not find a public database on this topic. please guide me.
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Dear Shafagat Mahmudova
Thank. I need the
article "Novel structural descriptors for automated coloncancer detection and grading" dataset.
Unfortunately, the author does not answer my emails.
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I am recently doing PDX model but find serious fibroblast cells contamination. Can anyone share experience on that one?
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Would you like to make 3D culture from colon cancer cells alone or from all the cells found in colon tumor microenvironment? Personally, I tried 3D Bioprinting method, it works very well in many conditions: http://www.n3dbio.com/products/magnetic-3d-bioprinting/
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As there is a lot of buzz about Immunotherapy, which is mainly antigen-based, In Dendritic Cell therapy or T cell therapy, we need to stimulate cells by a specific Tumor antigen.
My question is related to tumor microenvironment what percentage of cells expressing the antigen in the tumor?
Tumor microenvironment has its mechanisms to escape immune response in which antigen loss is also critical.
Why Can we not use direct antigen for tumor Immunotherapy, if we can then how effective it is?
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Dear Prashant,
Back to your original discussion, yes antigen loss and tumor micro-environment are both major roadblocks for immunotherapy. A good example is that thus far, direct antigen cancer vaccines targeting single tumor antigens (MAGE, WT1, NY-ESO1, have not shown dramatic therapeutic efficacy.  https://www.ncbi.nlm.nih.gov/pubmed/27058246
Even with engineered CAR T cells, it has been observed that antigen-negative relapse happens in about 30% of patients. https://www.jci.org/articles/view/87366
The reason why Checkpoint inhibitors work effectively in a subset of patients (along with several other reasons being studied) could be that checkpoint inhibition releases the brakes on a polyclonal immune response and therefore the immune system is targeting several antigens, not just the 1 or 2 that cancer vaccines use. This is suggested in observations where relapse after checkpoint inhibition occurs in tumors which have lost B2M and therefore have lost the ability to present antigen.  http://www.nejm.org/doi/full/10.1056/NEJMoa1604958#t=article
Therefore, certain cancer immunotherapies (CAR T cells, checkpoint blockade etc) have already proven to be effective in clinical trials, but relapse does occur due to antigen loss, and thus they can still be improved.
It will be interesting to see what clinical trials show in the next few years combining immunotherapies with other drugs that have synergistic mechanisms of actions.
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Hi, does anyone have an information regarding further treatment options for a 61 year old male with mCRC with secondaries to the left lung and mediastinal nodes who has DPG deficiency confirmed after two doses of Avastin and Folfox when he presented with neutropenic sepsis/typhilitis and local perforation. Any options other than single dose irinotecan?
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Hi, try with mitomycin C plus irinotecan, or with regorafenib or pemetrexed. You can find several phase II trials suporting this therapies. 
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Dear Researchers Working in field of Tumor immunotherapy,
I am working with MC-38 colon cancer models. I had faced a critical issue when I tried to use TAA immunotherapy.
I have observed this phenomenon twice:
To study the TH1 specific response we are considering IFNγ and TNFα, When I am culturing splenocytes of tumor-bearing mice overnight with Antigen, I found there is a loss of CD8 cells. Is it very common phenomenon or I am making some mistake?
Awaiting for your crucial comments
Thanking you
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Hello,
I worked on invariant natural killer T cells, and I found that these cells become anergic upon activation with a specific antigen and they down regulate their specific receptor. you may have the same situation with CD8 cells (anergy + down regulating their receptors). And this is not the case when isolating healthy NKT cells.
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Dear Tumor Immunology Experts I am trying to isolate Tumor-infiltrating lymphocytes primarily T cells from tumor-bearing mice induced by MC38 cells.
I do not have CD45 beads, but I am using CD45 antibody. I have collected total tumor cells and run the FACS, as you can see the SSC and FSC population in the figure I have attached:
No-1 Is the whole cell population with voltage adjustment to visualize 100 percent cells.
No-2 is also a whole cell population with voltage adjustment to visualize only lymphocytes.
When I go with no-1, the CD8 and CD 4 population percentage are tough to gate even I am using CD45+ cells, but when I am going with no.2, the CD8 and the CD4 population is very easy to distinguish.
It is better if I am using with a no. 2 of voltage, but I am not sure if it can be accepted for publication or not.
I am looking for the valuable advice and guidance.
Thanking you
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Dear Prashant,
I have some experience with immunophenotyping MC38 tumors ex vivo. 
The way I approach it is to make sure the larger part of all cells (tumor and immune) is in the FSC/SSC plot. Then I gate on alive CD45+ cells, then on CD3+ cells and then on CD8b and CD4. I end up with a pure population of T cells. Remember there are other CD8 and CD4 populations, for example dendritic cells, that are not T cells.
To answer your question; the most important issue is to make sure that you do not exclude your cell population of interest in any gating strategy. As such, I would advise to backgate your T cells to show there are none that fall out of the FSC/SSC gate. 
Personally, I do not see a problem in using your second approach (NO-2) as long as you make sure to not exclude any T cells (back gate to make sure).
One small thing, in your second plot, I would not advise to call the gate "lymphocytes" since that gate may contain many other cells.
Hope this helps
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Adjuvant radiation therapy is beneficial in T4 colon cancer in ascending and descending parts.
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Dear colleagues,
nowadays one of my friends who want to assay some genes and proteins in mice suffering from colon cancer. But i want to know that how he can become sure that his treated animals with high dosage of arsenic are a suitable model or not after treating? Does anybody knows some indices or landmarks as well as markers for checking animals to being ensure that his animals suffere from cancer?
This massage is wroten by cellphone.
regards.,
Mehdi
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Dear Nicholas,
Bench of thanks for your reply., do you have any experience in it?
Do you have any literature in this issue?
Regards,
Mehdi
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The work is related to my Ph.D work.
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Hello 
Do you think benzene would be helpful to induce cancer in the mice model??
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I am analyzing RNA-seq data from colon cancer samples using the RPKM method.During the analysis pipeline , the bed file that i created using bedTools does not show the gene ids.And also, the bed file seems to be larger than the sorted bam file(almost double).What could be the problem?
I used bowtie for alignment.I used hg38 refseq genes bed file as the annotation file.
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Check the Biostars answer at https://www.biostars.org/p/163292/
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I am using DMH to induce colorectal cancer with 20mg/kg dose twice a week for two weeks subcutaneous injection.. 
Please, refer me how to be sure that my injection is right and the accurate subcutaneous injection, and give me any helpful advice it is my first research.
thanks a lot
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DMH is the most widely used animal model for colon cancer and this chemical induces colon adenocarcinoma with highest incidence rate..
In general it is given as intraperitoneal injection for about 4 months followed by confirmation of colon cancer. 
A work by my colleague (link below) may be helpful to you. Evaluation of crypt foci is a major and useful parameter as well.
Best Regards,
Grandhi V Ramalingayya
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i m searching for papers that have an injection dose of stem cells that used to treat colon cancer ..
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i think using mesenchymal cells may enhance tumour progress if they act as immune supressants
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Anybody knows that when the adjuvant chemotherapy (ACT) of colon cancer was recommended for the first time in the guidelines?
The first evidence of the survival benefit of 5FU-based ACT over surgery alone in stage II-III colon cancer patients was reported from NSABP C-01 trial in 1988, which was afterward confirmed by later pooled analyses. Following these pioneering studies, 5FU deemed to be the standard control arm in newer CRTs which were consequently unable to investigate the effectiveness of the innovative surgery alone treatments (e.g. complete mesocolic excision) of colon cancer. Surprisingly, The disease-free and overall survival benefit of ACT reported in NSABP C-01 trial disappeared after ten years in an updated analysis, making a glass ceiling for the real effectiveness of ACT in clinical practice. I am working on this story and would like to know how the ACT came to the guidelines as a standard treatment of colon cancer.
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I think NSABP C-01 was the positive study showing some benefit from adjuvant chemotherapy:
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Please refer me any protocol for developing  colon cancer animal model in particular to mice and zebra fish where I can study metastasis.
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Why you prefer Zebra fish model for metastasis?
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How should I calculate the cut off for lymph node ratio in colon cancer, using a national data set?
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Dear Dietrich, Thanks for your reply and sharing your interesting paper.
Dear Bruno, Thank you too for your excellent answer.
I will back to you after applying your suggested methods in the analyses.
Best regards,
Masoud 
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is there any er stress staining method? 
and best for colon cancer cells?
I want an easier method to take a photo for er stress phenomenon.
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ER stress is a biochemical phenomenon that can be measured with biochemical parameters. I don't think photography is the best method to study it.
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I am looking for MC-38 (GFP/RFP) also known as colon-38 and SL4 cell lines for my study. If anyone have it in their lab, I would be happy to contact.
Note: We got the MC-38. Please ignore the question.
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I need to explore  EMT in colon cancer in vitro. I have read some papers written by other researchers,  they usually choose SW620,HCT116 .Someone said the morphology of them is different .But when I search on www.ATCC.org , I see the morphology of them is both epithelial.I want to ask  this is why.
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Hi Siming. The term "epithelial" is a bit of a "global" term and covers a number of cell types that may have different (sometimes subtle) morphologies.
In fact at its simplest there are only around 4 classification for cells: Fibroblast-like, epithelial, neuronal and lymphoblast-like.
Thus two cell types can be termed "epithelial" but look quite different. When I was looking into colon lines a few years ago I found the following paper (Ahamed et al 2013)  a great help (they show images of a number of different colon lines - including the two you are looking at if my memory serves me right.)
Hope this helps,
Gary
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What should be the endpoint (in the context of tumor size) of an experiment in which the animals are injected with CT26 cells? The idea is to measure the tumor until the endpoint, at which point the mouse bearing the CT26 tumor will be sacrificed? Generally, when we work with MC38 or B16, our endpoint is when the tumor size approaches 2cm x 2cm.
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Currently working with 5 cell lines, B16 and CT26 being two of them. For CT26 I once harvested the tumors for down stream analysis at sizes 15mm by 14mm because at that point mice overall health appeared compromised.
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I am looking to create a cell line stably expressing WT K-ras in easy to transfect cell lines like HEK293T or HeLa.  Cell lines like HeLa and 293 seem to be used predominantly in a transient transfection fashion, while there are some studies suggesting that K-ras over-expression in HeLa cells is not good for them.   I see that colon cancer cells are successfully used for stable WT K-ras and mutant K-ras - of course, K-ras is important for colon cancer studies.
Has anyone generated stable WT and mutant K-ras clones in Hela or HEK 293T? Are there any limitations to these cell lines? What other "generic" cells can be used?
Thanks.
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The choice of a cell line to stably express or overexpress K-Ras would in large part depend upon the question(s) you would like to ask using the cell line. Further, there are other considerations such as "overexpression" could lead to retardation of cell proliferation and elimination of "clones". Two cell lines that you mentioned are not good to study cell cycle regulation or cell growth regulation. One can consider cell lines, which are deficient in the K-Ras function and restoration of the function may be of interest.
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Hello everyone, 
I'm starting working with SW48 colon cancer cells in vivo, and I wondered if Matrigel is needed for a subcutaneous injection. And, how much cells do you generally inject? 
Thanks a lot for your help!!! 
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SW480 cell form tumors upon the injection i.p. of 2x10(6) cells within 2 weeks. If you plan the s.c. administration, 0.5-1.0x10(6) cells should be enough. You don't need the Matrigel, albeit it keeps the cells together and thus supports the tumor development. If you will use the Matrigel, the number of cells injected can be even lower. 
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 want to know how do you detrmain the necessary dose of silver nanoparticles to treatment colon cancer in mice?
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Are you functionalizing commercial colloidal silver? In that case I would recommend stirring them in aqueous NaBH4 solution with excess lipoic acid. Alternatively, you can reduce Ag+ directly with NaBH4 in the presence of lipoic acid. As far as determining the necessary cytotoxic dose, I suggest starting with a colon cancer cell line and treating the cells with varying concentrations of your nanoparticles. Then you can determine the cell viability by MTT assay.
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while  operative any cancer case the level of resection highly depend on metastasis of lymph node. however, micrometastases are not visible through nacked eye and our surgery goes blindly ! if we are able to determine the level of micrometastatic lymph nodes. I think we could increase DFS and OS in colorectal cancer.  so can any one suggest me other than Indian ink or carbon nanoparticle? wich can give a more precise result.
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Firstly thank you all of you, but i want to asses on operative table which must be less time consuming and give more exact idea about the presence of pathology . same as fresh frozen biopsy.
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67 old who had left Hemicolectomy for adeno carcinoma T3N0M0 followed by adjuvant chemo 15 Years ago, Now he is found to have recurrent cancer near the anastamosis on screening colonoscopy.Cect showed few enlarged pericolic nodes near the tumour.No distant mets.He is symtom free.CEA and PET reports are awaited.
.4years ago his daughter under went Left colectomy for the same.
would you recommend Total colectomy with Ileo Rectal or IPAA
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There is a correct protocol and a right decision. Good luck again!
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While left colectomy for cancer is widely performed by miniinvasive surgery, there are, to date, numerous surgical centers that still practice right hemicolectomy totally by open approach or only partially by laparoscopy (with variations depending on whether colic mobilization and/or vascular ligation are made laparoscopically or not). Only few centers perform right colectomy totally by laparoscopy (vascular ligation, colon mobilization, colon specimen extraction by enlarged trocar incision or Pfannenstiel mini-incision, intracorporeal ileo-colic anastomosis). What do you think about?
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If it will be easy to do mesocolic excision by laparoscope, Ok
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A 46 year-old patient, with performance status of 2, diagnosed with colorectal cancer underwent a partial colectomy ( R2 margin). The thoracic CT scan described infracentimetric secondary lesions in both lungs. The abdominal CT scan described multiple hepatic metastases (image attached).
My questions for you are (especially for the surgeons but also for medical oncologists):: 
1. do you think this patient could benefit from resection of liver metastases? Not even after chemotherapy+biological therapy?
2. what is your opinion about the surgical management of this patient?
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Dear Dr Afrasanie
Inorder to decide that patient can benefit from liver metastasectomy , we should have all liver plan and radiologist should say about future liver remnant (FLR) , it should be more than 20% in normal situation and at least 30% in patient who received chemotherapy.
Besides we should be sure about extra liver situation such as lung metastasis.if there are mets in lung it should be operable.
For better judgment about how many mets in liver or lung , new studies recommend PET CT scan.
However for yours patient , according one cut of liver , it seems to be at least borderline or inoperable . In optimist situation, neoadjuvant chemotherapy in orer to down size of tumor or left portal vein embolisation in order to hypertrophy of right liver lobe is an option to have a secure FLR.
Good Luck.
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While microtubule targeting agents are extremely effective in other forms of solid cancers, none of them is approved for colon cancer. Is there some specific form of resistance of colon cancer cells to these drugs?
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Thanks very much all for your input. The P-gp idea seems indeed like a good explanation. Colorectal mucosa does express high levels of P-gp (see: http://www.ncbi.nlm.nih.gov/pubmed/7577038).  However, patupilone (epothilone B), which is not a substrate for P-gp, also recently failed Phase 2 trial in patients with colorectal cancer (see: http://www.cancerresearchuk.org/about-cancer/find-a-clinical-trial/a-trial-looking-at-patupilone-for-people-with-advanced-bowel-cancer).
Perhaps it is a combination of P-gp with some specific biochemistry in the actual colorectal tumors, which cannot be easily identified (or absent) in in vitro colon cancer cell cultures. Perhaps this is the reason for the absence (to my knowledge at least) of literature on this topic. As Robert Kiss (who responded to this question earlier) taught me, tumor environment plays a key role in drug resistance.
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Hi friends,
I am trying to determine ROS by FACS analysis.
The protocol which I followed here,
In this experiment I used colon cancer cell (HCT116)
I used as a positive control H2O2 treated cells (100uM, 1 h treatment)
1. After treatment the cell were washed with PBS and incubated with 30uM 2’,7’ –dichlorofluorescin diacetate (DCFDA-Sigma) for 30 mins.
2. After incubation the cell were washed  with PBS and trypsinised and collected in PBS and keep in ice until FACS analysis.
After staining with DCFDA, When I try to acquire the data by FACS, I am getting strange data.
Usually, H2O2 treated cell should shift to right side, for me in contrast it shifting to left side. I dont know why its happening for me?
I have attached image with all condition of FACS setting.
Please find the attachment.
Hope to get some positive reply from every one.
why it shifting to left side?
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Hi Pasupathi
You should increase your H2O2 exposure times. Try a kinetic of H2O2 (2h, 4h and 6h). You can and should also titrate H2O2 concentration (100, 200 and 400uM).
As for the flow cytometry analysis you should
  • eliminate doublets by a FSC-H vs FSC-A (you need a single cell analysis and during flow acquisition more than one cell can pass through the laser).
  • gate on live viable cells using PI or 7AAD negative population (H2O2 can induce apoptosis and dead cells have autofluorescence)
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Hello,
We have a colon cancer cell line that has been for us for many years and it shows different behaviors than it originally should do. When we did a literature search, we suspected that the cell line could have changed its genotype over the years and we thought that we should do an STR (Short Tandem Repeat) analysis to authenticate the cell line's identity. I found some loci and primers for those loci which are used for the STR analysis of the cell line but I could not find a suitable protocol nor do I know if it can be done in the lab with just the use of conventional PCR.
I am also aware of the presence of the Powerplex kit but we could not do it since we don't have the fluorescent imaging of PCR products in our department.
Do you know any related protocol or anything that might be useful?
Thank you all in advance. Enjoy your experiments! :)
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Hi Caner Yener,
I know that the topic is old, but out of curiosity have you been able to find out if the cell line is really contaminated?
Computationally speaking, RNA-Seq data of MKN28 cell lines always show pure allele of mutated TP53 gene indicating that all the cells have a single origin.
Thanks :)
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How would you treat a 55 year-old patient, with PS=1, and these features?
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Dear Dr Vlad-Adrian
please let me know the haptic lesion size and is it operable or no?
if hepatic lesion is operable and it has a notable size i prefer to start with neoadjuvant chemotherapy such as FOLFOX or FOLFRI with AVASTIN. Before the disappearance of liver mass follow chemotherapy in control CT scan , i will operate in one or two stage and finally if optimal surgery is possible, i recommend HIPEC.
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I want  to study protein-protein interaction between integrin beta1 and EGFR in cell membrane and cytosolic fraction of HCT116 colon cancer cell line.
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Wael Thank you. I realized that i was not using enough antibody to pull down the protein. Thanks for your advice.
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a 55 yrs old woman with a past history of colon cancer that underwent left hemicolectomy ,T2N0 well diff adenocarcinoma, in follow up colonoscopy one year after operation shows a 1*1 cm cessile polyp that removed endoscopically . pathology showed hi grade dysplasia .
what is your recommendation?
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Dear Dr Negar
Thanks for your attention. according NCCN ver 2,2016 for sessile polyp ,two option was recommended but regard to patient history of cancer i am keen to surgery.