Questions related to Cancer Cells
When I isolate RNA from cultured cancer cells and check for different epithelial (E Cadherin, CK19, Klf5)and mesenchymal (Snail, Twist, Zeb1, Vimentin etc) markers by qPCR, somehow the trend of expression of these markers is never consistent. Does this happen? Is there any reason?
Sorry to bother in this Sunday
But I would like to ask a few questions:
Does anyone know about how to induce the generation of polyploid giant cancer cells (PGCCs) in melanomas with "targeted drugs"?
How to measure the presence of polyploidy in the tumor?
How to induce the generation of giant polyploid cancer cells (PGCCs) in melanomas?
Is there any protocol to induce the generation of polyploid giant cancer cells (PGCCs) in melanomas?
Is there any way to study the immortalization and regeneration of tumors with cellular and molecular biology? Is there any protocol?
Matheus Correia Casotti
I am treating some cancer cells to see if they are directly linked to centrosomes. I did immunostains on gamma-tubulin, flow on cell population after treatments, and qpcr on some centrosome-related genes. I am just wondering what other methods I can use to show that my drug or particular active ingredients affect cell centrosomes. Thanks.
In the previous MTT assay, it was confirmed that the survival rate was decreased, and in the DAPI experiment, an increase in the apoptotic body of the substance-treated group was confirmed.
After that, it was confirmed that there was more blank space as the cell viability decreased in the substance-treated group by treating the cancer cells with a substance.
However, as you can see, the cleaved-PARP, which should be increased, came out more strongly in the control, and also the bax decreased compared to the control. Have you ever had a case like this?
* The top line is total .
Nano-medicines are widely used in Oncology but are their actions on specific target cancer cells or they act on nirman cells also?
I want to ask you because the morphology of the HSC-3 cells I grow is different from the general shape. If the cells look like this, what's the problem?
I am trying to do ATAC seq for my epigenetic research on human prostate cancer cells ( PC3 and LNCaP). I have a bottle of genomic lysis buffer. I am not sure if I can use it for ATAC seq. I would appreciate anyone answering me or suggesting me the right buffer. Thank you.
I'm doing apoptosis assay using the flow cytometer on cancer cells (mcf-7). I'm wondering what's the ideal event run count and speed to be used?
Can i used the commet assay to differentiate between the cancer and normal cell?
how commet assay result i interpret to distinguish that this is the normal cell and this is cancerous cell?
I am trying to figure out through what means Acetoacetate may be converted to acetate naturally within a healthy, or cancerous cell (though specifically non-hepatic cells). I am especially interested in any methods which involve the TCA cycle. I have been currently considering acetoacetate converting to acetyl phosphate then acetyl phosphate to acetate within the mitochondria; however, I do not know if this is a possible pathway. Any information upon potential pathways for this conversion, or how viable the acetyl phosphate pathway is would be greatly appreciated.
Hello, I am reading a paper saying their compound can lead to apoptosis of HepG2 cells; however, it came to me that is it possible that the cytotoxicity of the compound causes the apoptosis? Or what’s the difference between cell cytotoxicity and cell apoptosis?
Let’s say, If one compound is capable of leading to cancer cell apoptosis, how about its function on normal cells? Can it cause the apoptosis of the normal cell too?
I got an experimental result where the mitochondrial transmembrane potential of drug-treated cancer cells has increased than untreated cells, as well as ROS generation, was found to be higher in treated cells. The experiment has been repeated multiple times and a similar result was found. The experiment was based on JC-1 and rhodamine-123 staining method.
Thanking you in anticipation for your answer and suggestion.
on a tight budget with little experience in studying cancer in mice which method should we pursue for successful implatation of MCF-7 cancer cells on mice
I am now conducting research on HER2-positive and HER2-negative breast cancer cell lines. I'm looking to confirm the overexpression of HER2 receptors in HER2-positive cells versus HER2-negative cells. Could you possibly assist me in determining how I may accomplish this either quantitatively or qualitatively?
Im going to plan a cancer study. Im appriciate to read and answer my question.
1. Using NCI-N87 gastric cancer cells, i'll induce subcutaneous tumor in mouse.
But between flank and hind limb, i cant choose the region for cell injection.
Which one is better between them?
2. Im also wonder the necrosis in growing tumor tissue. i want to take MRI imagies and tumor vascular histologies. How can i take care of the tumor mass to prevent necrosis?
It is an experiment to check the effect of a certain phytoestrogen on a particular cancer cell proliferation in-vitro.
The results consists of
Six absorption readings from control group
Six absorption readings from X phytoestrogen at concentration A
Six absorption readings from Y phytoestrogen at concentration a
Six absorption readings from Y phytoestrogen at concentration b
Six absorption readings from Y phytoestrogen at concentration c
I am new to culture HUVEC in 3D collagen on a microfluidic chip. I recently met a problem.
The problem is that the HUVECs will die after being cultured in 3D collagen for 2 days. in addition, A549 lung cancer cells also die when culturing for 3 or 4 days. In fact, the cells are seeded onto the surface of collagen lumen. However, the cells remained in inlets/oulets and PDMS channels live well.
Therefore, I think the reason may be the shortage of nutrients for cells in lumen.
I have no one to ask so I want to get some help from you guys.
any suggestion is welcome.
I am looking for a method to assess the function of cancer cells and macrophage co-culture. rather than inducing the macrophage to either only M1 or M2, I would like to see what is the effect of coculture between a spectrum of macrophage and the cancer cell in vitro. I think this can recapitulate more the in vivo microenvironment where both M1 and M2 macrophages co-exist.
I am using both the J774 and THP-1 cell lines. May I know if there is some well-established protocol/ published article that tried inducing the differentiation of both M1 and M2 at the same time?
Articles remark that cells plated at the right density have enhanced efficiency of organoid culture establishment. Plating too densely will result in increased cell death at the core of the dome.
As my cancer cells have undergone quite amount of passaging , thus with every passage , there are stress factors which either float around them or are stuck on the cells, as a result my cells are not geting the growth media , i have tried everything by filtering the media, increasing the antibiotic concertration and also lowering FBS but the stress factors are still present , i have been changing their growth media in every 12 hours now , is there any other way to make them go back .Please share your view
Looking for a fast-track journal to publish one of my papers in the field of image processing for non-hodgkin leukemia cancer cells detection method.
What about irradiate nanoparticles as a metal like ZnO or Iron oxide to treat cancer cell.
Any suggestions about the techniques of the exposure and the dose that kills 50% of cells
We have been using the Celigo to count and measure the size of breast cancer cell mammospheres and we are having issues getting accurate results. The machine tends to break up large mammospheres into 3-4 pieces and count them individually, while also counting a bunch of smaller individual mammospheres as 1, and also counting things that are just single cells near each other.
I have played with the settings numerous times and have worked with a technical person from Nexcelom and cannot get the settings to work for us. I was wondering if anyone had any luck using the Celigo for these purposes? I have tried analyzing the mammospheres using the Colony setting and the Tumorsphere setting.
Thank you in advance for any insight.
I am doing the NO assays using griess reagents without kit, please tell me what will be working concentration of griess reagents and how the can be done spectrophotometrically. Thanks
TGF-β and BMP signaling are present in the cancerous cell show almost the same function of chemoresistivity, enhanced proliferation, higher EMT transition. Then why would cells have both mechanisms? Also, what is the major difference between both of them?
I'm trying to understand the mechanism of L-amino acid oxidases (LAAO) on cancer cells and healthy/normal cells. Why can LAAO treatment induce apoptosis efficiently with a low dosage on cancer cells compared to healthy cells? Can you guys help me with clarification on this problem?
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.
Good Day I don't know if you guys will read these but I really need an advice.
So here's the thing I compiled all the authors available from various databases about the anticancer property of Peperomia against multiple cancer cell lines.
These are the sample data that I got from multiple papers
Roots > LC50 > against various cancer cells (including cell viablity)
Leaves > LC50 > against various cancer cells (including cell viablity)
Stem > LC50 > against various cancer cells (including cell viablity)
Roots > LC50 > against various cancer cells (including cell viablity)
Leaves > LC50 > against various cancer cells (including cell viablity)
Stem > LC50 > against various cancer cells (including cell viablity)
What would be a good way to represent my data in my paper? Is there any statistical tool or analysis that I can use?
Thank you sincerely!
We are planning to use Anti-Cisplatin modified DNA antibody for fluorescent microscopic imaging of cancer cells. In this regard, availability of specific protocols for staining cells with such antibodies is required.
Suggestions are highly solicited.
We know that the brain sends and directs meaningful messages to control the patient's cells.
as we know, The brain is affected by factors such as diseases And we know that the brain also controls other organs of the body.nevertheless,Damage to the CELLS is visible on eeg?
Is Cancer Effective In EEG?
The dielectric constant of cancer cells has been obtained experimentally and has been stated in some articles at a certain wavelength. Is there a report on the refractive index of cancer cells, especially mcf-7 cell, at different wavelengths?
I need this very much because I have no experience on it. So I want to publish an article by using this kind of web based tools.
There are many method to check about stemness of cells. But, I need a method that can compare stemness within few minutes. Is there any method ?
Can we perform spheroid formation and invasion assay of cancer cells using Lionheart FX Automated Microscope?
when designing a chimeric antigen receptor for cart t cell therapy can't we use a target which is also exist in T cells but express excessively in cancerous cells?
I want to do some cell migration assays and I would like to know if it's possible to grow these cells in suspension
I'm studying about cancer cell and its pathways and here I have some questions. Do cancer cell still have their wild-type proteins or genes? or all of those wild-type proteins and genes already turned into mutant ?
any answer will help me a lot!
Thanks in advance!
I wanted to buy an incubator in which we will be maintaining neuronal as well as cancer cells. With advancement in technologies, i was wondering whether air or water jacketed incubator is best for academic lab purposes? All your inputs are much appreciated. Thanks
I was performing a coculture experiment of cancer cells along with CAFs and my goal was to look for the CCND1 gene expression (Hypothesised to go up in the coculture).
What I observed was that the uncocultured cells showed higher CCND1 expression compared to the cocultured ones.
Is this due to an effect observed due to high confluency of cells leading to contact inhibition and decrease in CCND1 level?
Appreciate any kind of output!
Hello, I am trying to isolate single cell colonies form MCF-7 breast cancer cells. I didn"t get any colony in my plate. they were edited by CRISPR but the deletion does not affect their growth/survival. Thank you!
I am using a lentiviral TET-ON system for conditional overexpression of my gene of interest in primary kidney cancer cells. I use viral load from transfection of HEK293T cells after 48 and 72 hours and perform selection for 2 weeks using Puromycin (using a concentration I have optimized for the cells I am using). After to weeks, I use Doxycycline at 2mg/ml (adding fresh media everyday) for 72 hrs. However, in western blots, I only see slight increase in expression of my gene of interest compared to -Dox. How can I increase the efficiency of overexpression?
For the evaluation of Lysosomal destabilization in cancer cells, if there are two different drugs encasulated in NPs then how the effect of NPs containing two different drugs and their combination can be evaluated using Lysotracker red DND. If the two different NPs cause lysosomal destabilization in their individual treatment then the combination of both the NPs should cause impressive result like more lysosomal destabilization.
Then if the combination shows more lysosomal destabilization then what would the impact i.e. towards which phenomena it indicate or if lysosomal destabilization in a lesser fraction then wich phenomena will it indicate? Does it has any relation with autophagic flux?
The entire process can be evaluated by laser confocal microscpopy?
What might be the possible reason for granulation of ct26 cancer cell line?
I tested media(RPMI), serum and all of cell culture conditions but I cant get rid of granular cells.
After collecting the cancer cells treated with anticancer substances, the centrifuge is rotated at 1300rpm for 30 seconds, and after removing the remaining pbs, 1*10^6/80ul LYSIS BUFFER is added based on the number of cells calculated. And (voltexing 1 minute, freezer 5 minutes) After repeating this process 4 times, it is rotated to 1300 centrifuge for 5 minutes, and the supernatant is collected and used as a Western protein. c parp shows a good trend, so apop seems to be concentration-dependent, but bax and bcl2 tend not to show well. The desired result is an increase in bax and a decrease in bcl2. What conditions would you like to change?
Is that CCK-8 test can also react with DH5a (E.coli)?
In numerous studies have shown that CCK-8 or MTT tests for detecting the proliferation of cancer cells incubated with certain bacteria. However, in my recent work, I find that DH5a also presents a great influence on the OD450 of the CCK-8 test. I really wonder the validity that using this method for examining the proliferation of co-culture bacteria and cancer cells. Furthermore, I apply Hematocyte Counter for checking this result, I find that the so-called phenomenon of cancer cells proliferation have been definitely disappeared, or even displayed cells number decreasing. I did not know how to explain these conflicting results.
Many thanks for your kindly response.
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?
I am using JC-10 (Enzo life science) dye for quantify mitochondrial membrane potential in CRC with 10 uM final concentration. I treated cell with CCCP 10 uM for 20 min for a positive control. I took the reading for J-aggregate at Ex/Em: 490/525 nm and for monomeric JC-10 Ex/Em :490/590. However I could not get the appropriated 525/590 nm ratiometric decrease in MMP even in CCCP treated set. I also repeated the experiment applying JC-10 in serum free medium to stain the cell, and again I Couldn’t get the result. I have a doubt in my protocol of staining buffer. Many companies supplying JC-10 kit provide two types of assay buffer to stain the JC-10 in the cell. I could not find the information about the ingredients mixed in such assay buffers.
If you have any suggestion for me in staining buffer and in whole procedure. Please help
Hi all. I would appreciate any help regarding this. I am trying to grow patient derived primary bladder cancer cells from surgical samples using mechanical and enzymatic digestion approach. Initially cells were growing well but after few days, it seems the cells go into senescence stage. I have attached the corresponding images of the cells after 21 days. Any suggestions/comments on this would be very helpful.
Currently I am using DMEM(high glucose)-F12 media with 3% FBS + EGF-insulin- Cholera toxin-hydrocortisone as supplements.
Thanks and regards
I have been doing some experiments to reveal cytotoxic/antiproliferative effects of the new compounds in the cancer cell lines. After 24 hr incubation with compounds in Hep3B cells, I realized a specific structure on the wells with different compounds even my positive control (adriamycin), but not on control wells. So I am attaching an image of the wellhole shaped structure. Can you give an idea what this structure is or looks like? Thank you
I want to measure the cellular uptake of nanoparticles in cancer cells using ICP-AES. I want to know the concentrations of NP (subtoxic concentrations or high concentrations) and the incubation of NP with cancer cells ( short time or long time) and the protocol for cancer cell digestion before applying to metal analysis using ICP-AES measurements.
Thanks in advance.
My lab team and I are planning on making a co-culture with mesenchymal cells and cancer cells. The thing is that mesenchymal cells were grown in DMEM, and according to the ATCC the cancer cells are supposed to be culture in RPMI. Is there any protocol that can help us determine how to adapt the cancer cells to DMEM in order to make the co-culture?
Any suggestions would be appreciated
I am looking for breast cancer cell line models that reflect the PAM50 HER2-enriched (HER2-E) classification, which may overlap, but not be necessarily the same as HER2+ breast cancer cells.
5-fluorouracil, oxaliplatin and irinotecan purchased from Sigma are failing to show an inhibitory effect on cancer cells (tested on HT29 and A549) in cell viability assay (MTT). Please advise on what could have gone wrong. They are not expired. I have tried preparing fresh solution as well-also did not work.
Please note that I am well-versed with MTT assays and have in the past worked extensively with 5-FU and other chemotherapeutic compounds, so I do not expect it to be a problem with a standard assay such as MTT.
Thank you for your help in advance!
I am currently writing an introduction to my thesis and reading "Biology of Cancer" of Weinberg.
The book is so awesome and interesting, I just can not express how much I love it: it touches all themes of cancer, starting from "what is a cancer cell" and finishing with the biological points of its development/biology (nearly 1 000 pages).
I would like to write an intro to my thesis which would be a pleasure to read, and I would like to find an inspiration in the book, like the one about cancer. I don`t want reviews (already downloaded them), I would like scientific book, which would cover many aspects of telomeres and/or aging.
Could you recommend one, please?
Thank you very much for all suggestions.
Soon I will be making a co-culture of cancer cells with mesenchymal cells. I have cultured the mesenchymal cells in DMEM for several weeks and they have responded fine, but I do not lnow if it is suitable for cancer cells. I do not have much experience working with animal cell culture, so any feedback is very welcome!
Thank you in advance!
Best wishes from Costa Rica,
I am currently working on a project that involves characterizing FT190 and FT194 cell lines, but I can't find much information about them. I want to know as much as I can about them, including the patient's nationality from which they were derived from and if they are chemo and radiation naive. Thanks!
Recently, I am exploring the potential function of one gene in cancer. Since the TCGA datasets as well as some other clinical data show that this gene is up-regulated in cancer and especially in basal-like subtype. I tried to knockdown it by shRNA and the result show the depletion of this gene accelerate the proliferation, migration and invasion rate of cancer cells. I also tested the over-expression in cancer cells and the result show the over-expression impaired the proliferation rate. This observations is also consistent with some previous report which reported it as tumor suppressor. Although I found that expression of this gene maintain the basal-like feature of breast cancer. The results is still confused me.
My question is whether you know some similar works or papers.
Dear esteemed researchers,
I am looking for proteins from breast cancer cell line MCF-7 to study docking of my proposed organic compounds. Please guide me about the most significant proteins from MCF-7 which should be used to study docking interaction of organic compounds.
I am hoping to identify the proteins present in different areas of primary cancer cells (cytoplasm, nucleus, membrane etc.) by mass spectrometry. However, my access to fresh cells is limited, therefore I am hoping to carry out this experiment on frozen primary cells. The ProteoExtract Subcellular Proteome Extraction kit looks promising and is compatible with frozen cell pellets. But I am finding it difficult to find research papers that used this technique to analyze frozen primary cells and not cell lines. If anyone has any experience in this area, I would greatly appreciate some guidance? Thank you!
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
Nowadays i make recombinant protein. but i can`t see single band. alwasys i see multiband in E.coli system (in reducing system but multiband). but when I overexpressed that protein in cancer cells, I always see single band (this time I use myc or flag tag)
his tag problem?? or just expression species problem?
I am working on oral cancer cell line and i would like to observe the in vitro antioxidant activity in treated cancer cell line to evaluate the enzymatic activity like SOD, CAT, GST. I have read many papers and they have tried inducing oxidative stress in the cancer cells to observe this activity. I would like to know why we need to induce it when the cancer cells can also produce free radicals.