Questions related to Cancer Metastasis
...For research purposes to help others. If you are or know of anyone who is who would be willing to share his/her story, I would greatly appreciate the introduction! Thank you so much!!
I am a newby on in vivo imaging. We are using IVIS imaging system to study breast cancer metastasis in vivo, but we really do not know how to quantify the images along time. As the time pass, of course, the primary tumor and metastases grow so we must change the adquisition parameters. I would like to know how to properly quantify to able to compare the different time points. Thanks a lot!.
I need an ID8 murine ovarian cancer cell line to study the peritoneal metastasis in C57BL/6 mice. To perform noninvasive imaging of cancer metastasis, I also need ID8 cell lines expressing either luciferase or GFP.
Please kindly let me if anyone can provide me the cell line. Thank you so much for your support.
I am looking for an easy way to predict breast cancer metastasis through CTCs from whole blood or serum. Any idea? methods? targets? THANK YOU
I am doing a cancer metastasis experiment both primary and experimental metastasis. I would like to know has anyone collected CTC from mice blood and how did you store it?
I am looking to develop a model for lung cancer metastasis in C57 mice. I used a tail vein injection method but the model did not work for me. Do you think an injection of saphenous vein can help me? Or should I look for another way? Thanks a lot
I tried with Crystal violet but was not able to stain the colonies. It seemed that the stain could not penetrate the semisolid agar.
This is about a different approach to fighting cancer basically cancer cells must have glucose to survive. Following a ketogenic diet, the glucose drops and then using the administration of diabetic drugs to drop and keep blood glucose levels below 60 mg the healing process should start.
I would like to ask if you know people who tried this diet approach as detailed as possible & in the first place can somehow affect the patient they are any circumstances for that?
Can be this a way to heel cancer ? Are there researches that confirm this process or is the book written without evidence and is everything in theory?
I am planning to do orthotopic injection of breast cancer cells (MDA-MB-231 and 4T1 cells) into the mammary fat pad of mice to observe cancer metastasis and tumor growth. In lots of references I see people used growth factor reduced (GFR) matrigel and mixed it with the cells. My question is what is the purpose of using GFR matrigel. Will it makes a different when using matrigel containing growth factor?
Sometimes this can be accomplished by surgery, but the propensity of cancers to invade adjacent tissue or to spread to distant sites by microscopic metastasis often limits its effectiveness; and chemotherapy and radiotherapy can have a negative effect on normal cells
What do you think is the best way to treat cancer? do you know the side effects of the methods?
Do you have any information about methods such as radiotherapy?
Have you read an article about this?
Is it possible to make and xenograft a subcutaneous tumor with MDA-MB-231, and then induce a gene knockout in the tumor in order to analyze relevance of this gene in metastasis. Which system of inducible knockout could I use?
Australia's National Health and Medical Research Council (NHMRC) produced a sham review of Water Fluoridation in 2017 by deliberately ignoring over 3000 peer-reviewed scientific papers on Fluoride Toxicology. One important paper the NHMRC suppressed dealt with the mechanism by which Fluoride damages your teeth, with or without metal ions like Aluminium. Fluoride causes increased SATB1, a factor associated with Malignant Cancers and their Metastasis, with many relevant publications relating to Leukemia, Melanoma, Laryngeal and Nasopharyngeal squamous cell carcinoma, cancers of the Bladder, Breast, Cervix, Colon, Kidney, Lung, Ovary, Prostate, Uterus, and Liver.
A couple of papers: Zhang Y, Kim JY, Horst O, Nakano Y, Zhu L, Radlanski RJ, Ho S, Den Besten PK - "Fluorosed mouse ameloblasts have increased SATB1 retention and Gαq activity" PLoS One 9(8):e103994
Fluoride interferes with FoxP3
Zhang G, Zhou B, Han T, Wang M, Du X, Li Q, Wang J. 2012. Decreased percentages of CD4+CD25+ regulatory t cells and foxp3 expression in the spleen of female mice exposed to fluoride. Fluoride 45(4)357-364.
Observed increases in cancer incidence over four decades follow the roll out of Fluoridation and increased Dental Fluorosis. Surely it is time to ban Fluoridation worldwide?
What will be the best test in order to differentiate between Seminoma and Non Seminoma ?
a lot of data show that CXCL12 can promoter cancer cell migration
1. coating matricgel(1:9 in seriumfreeDMEM) transwell-chamber (SPLinsert 8 um) put into the Cell Culture Incubator 6hours
2. count the cell 2*105 use the non-seruim DMEM wash twice
3. inside chamber add A549 cell combine CXCL12 cytokine (100 ng/mL)
outside chamber add 10% DMEM
4. put in bake Cell Culture Incubator 14 hours than fixing and staining
this is my problem "CXCL12 treatment migrated cell is lower than the control"
I am working with antimetastatic compounds and would like to compare their activity with some other compounds. Maybe somebody know any approved drug from cancer metastasis? Or maybe somebody know good positive controls for migration and invasion assays in vitro or for both of them?
Three Days ago, the world lost a young and brilliant mind Maryam Mirzakhani (2014 Fields Medal, mathematics), died from metastatic cancer.
There are different methods which are under development, but why todays science can't be able to treat or delay the metastatic cells, I would like to ask which research line would be more promising in treating at least major types of cancer?
Hi. I am studying the role of exosomes in cancer metastasis. Exosomes derived from cultured tumor cells could be isolated and was identified by various methods. I was wondering if it would be possible to increase or decrease a specific microRNA inside the isolated exosomes in vitro? The plan is to use the microRNA modified exosomes to investigate the downstream effects or mechanisms if that could be done. Thanks a lot.
I am looking for Lung metastatic model for C57B6J mice. We inject LLC or B16F10 tail vein for experimental metastasis, but i want a model that tumor cell lines implanted in subcutaneous or intradermal so that metastasis can happen after several days. Suggestions are greatly accepted.
Injecting C57Bl/6 mice with B16-F10 cells via s.c. and i.v. routes. After primary tumor growth (in s.c.study) and pulmonary colonization (in i.v. study) we want to discriminate and isolate B16-F10 cells from the surrounding tissue, invading immune cells, etc. Is there a specific antibody (FC validated) that could selectively label the B16 melanoma cells? This would keep us from having to label and gate out the various subsets of cells found in tumor tissues (stromal cells, immune cells, etc). Any help would be appreciated.
We have been trying to develop 4T1 breast tumor model in balb/c mice. We have been injected 10^6 4T1 cells suspended in 100 uL PBS into the mammary fad pad region, subcutaneously. After 10 days we see mass, in site of injection. The size of mass is approximately 5mm ×5 mm. After 20-days mass is completely regressed in some mice and in some other the size of mass aggressively diminished. in pathological examination, some of the small mass have tumor properties and are malignant. if these mass are tumor what are the causes of restricted or abolished growth of them? why these cell line can induce small tumors in some balb/c mice but not in all of them?
The metabolism of most tumor cells is essentially higher than it usually is in most somatic cells. It is reasonable to assume that the tumor cell is much more sensitive to incomes of oxygen and carbohydrates. So, under blocking of feeding arteries, tumor cells should die first. The time needed for this could be set experimentally. So, if the local circulation is recovered after this blockade, we can expect that most of the normal cells will recover too.
Certainly, in the case of not local tumors, special procedures have to provide brain circulation.
Can Chemo drug elicit abscopal effect ?
Lysing (In Vitro) surgically excision tumor and admitting it back to the same patient who has multiple metastasis , will elicit abscopal effect ?
when cancer cells are inoculated in mouse circulation they will be immediately coated with fibrinogen ... in which it mediates there entrance into the different metastatic sites among them is the lung ... now cancer cells associated protein inside the lung willbe in which form fibrinogen or fibrin
i would like to see the fibrin that is associated with metastatic cancer cells and i would like to know which is the best stain that could be used to stain fibrin
brain cancer metastases from breast cancer
treatment of secondary brain cancer
We know that postoperative adhesions limit the availability and distribution of intraperitoneal chemotherapeutic agents to all areas of peritoneal cavity. It has also been shown that fibrin traps tumor cells and these cells in return become protected from any form of chemotherapy. Still the data on early postoperative intraperitoneal chemotherapy shows improvement in outcomes, especially for ovarian cancer. What is the mechanism of effectiveness if we know the drug is not evenly distributed in the peritoneal cavity and is likely not reaching all tumor cells.
I am working on the effect of fenofibrate on the 4T1 breast cancer metastasis by using Balb/c mice as the animal model ... but I have a problem in how I could give the required dose of fenofibrate without harming mice since most fenofibrate solvents will be toxic to them ... and the main problem is the limited solubility of fenofibrate in its various solvents like (DMSO, DMF and ethanol), so that to reach the required dose of the fenofibrate ( 0.2 % Wt/Wt), we will need a high volume of the solvent in which it will exceed the lethal dose .. also if we dilute the solvent we will need to inject mice with a higher volume that will exceed the intraperitonial volume dose ... SO I would like to ask kindly : 1 _ how we can give the required fenofibrate dose to mice without affecting them by the solvent ??? 2_ if we want to give the drug by food how we can avoid the variation in the amount of food intake daily by mice ??? Your time to answer my question will be appreciated ^_^ Montaser Haddad
Your time to answer my question is really appreciated ^_^
I am trying to isolate single cells from mouse lung with B16F10 metastatic tumours. Typically, after enzymatic digestion, I filter the cells through 70um filter, followed by 40% Percoll to remove dead cells. Finally, I lyse RBCs using ACK lysis buffer and proceed to staining.
However, as soon as I begin to stain (surface and intracellular markers), the cells begin to clump heavily and I don't seem to get enough cells for flow cytometry in the end.
I am unable to identify the reason for the clumping. Is it because of the tumour cells sticking around or because of too many dead cells? - I see about 40% dead cells after trypan blue staining, even after the 40% percoll step (~3ml percoll for 10min)
Also, I want to know if
1) Using a 40/80 Percoll gradient to separate tumour cells would help. I don't want to pool the lungs from different mice together, therefore is it advisable to layer the cells from one mouse only? I'm scared of loosing too many cells during this step.
2) Mostly I can get a maximum of 3 X 10^6 single cells from one lung. Is that what is usually seen or do people get more cells?
3) Is it advisable to pass the cells through a 30um filter instead of 70um? This is assuming that the lymphocytes (my pop of interest) can pass through it while tumour cells can't.
Any help would be greatly appreciated. Thank you !
Is metastasis something that every cancer cell can and will do at some point or are there genetic mutations or epigenetic changes are results in metastasis?
Does anyone know if E0771 from C57BL/6 can metastasize to bone regularly? Or, are there other murine breast cancer cell lines from C57BL/6 background that give rise to bone metastasis easily?
Thanks for your help,
I see many papers doing cell migration assay and then testing metastasis in animal model. Isn't metastasis inside body rather than cell based assay. Do we get any information or hint about metastasis from cell based assay?
It appears to be often quoted in various papers and articles although I've not been able to find the original source (papers often reference secondary sources only) - any ideas?
Among the array of cytoskeletal remodelling processes that a cell undergoes, surface phenomena such as protrusion formation (during migration and phagocytosis) demand an intricate coordination between the various molecular players involved. The significance of protrusion formation in cohort cell migration can be placed in a higher league. Considering its decisive role and the plethora of molecular players regulating the protrusion dynamics, what are the various parameters (biological and non-biological) that determine a stable cell protrusion
Hi there my friends, I have 4 chemotherapeutic agents to treat cancer cells. However, I'm not sure what is the best way to give the treatment: whether the 4 drugs have to be given at the same time in the culture medium or separately otherwise. Please help me and thank you all very much in advanced .
The role of autophagy in cancer metastasis seems to be complex. During early stages autophagy can play an inhibitory effect on metastasis. However, during advanced stages, autophagy acts as metastatasis promoter.
A recent review (Su et al, 2015) summarizes some of these aspects (see link). I am working in hepatocellular carcinoma, but my experience analyzing the relationship between autophagy and metastasis is limited.
Certainly many carcinomas (epithelial derived tumors) spread at least in part through the lymphatic system since lymph node metastases exist, and also are part of clinical prognostication. That in itself does not rule out direct dissemination to the blood. Is there any data on the relative contribution of lymphatic spread and direct blood spread?
Since lymph node (local) metastases have prognostic value one would expect them to be part of the mechanism of disease, i.e. there would be a mechanistic progression from lymph node metastases to diatant metastases. Is this born out by clinical observation? Can it, and has it, be mimicked in animal models?
We are looking for a mouse tumor cell line to do local tumor control study. We used 4T1 with small initial size (100-200mm^3) before. In a new project, we are going to increase the size to 1000 mm^3 when to start treatment and know it will take about >14 days. Because 4T1 has higher metastatic tendency and will die due to metastasis after inoculation, 4T1 is apparently not a good model for our experiment. Our research aims to develop a new therapy for breast cancer, so we have to find a breast cancer cell line which can grow in mouse with intact immune system (not incompetent one, like nude mice). Thanks
we know that EMT is critical step for metastasis ,then these metastatic cell require MET in establishment and stabilization of distant metastases. but what about cell lines taken from metastatic site , will they undergo EMT when inject intravenously and thus require MET in order to colonize metastatic site.?
I am studying the mechanism of a Serine/Threonine kinase to promote cancer metastasis. It will promote metastasis of cancer cells in vivo. But I can't find the target of the kinase to promote metastasis in vitro. So I am wondering whether I can use IP to get its whole substrates and then do mass spec.
Babak Rashidi et al. (2000) demonstrate that liver metastases from colon cancer are capable of remetastasizing to other sites ,1- however are there other tumors may follow this course ?
2- If metastatic cells re – metastasize , Do they Follow the approach that's similar to the primary tumor or that of metastatic site (as they originate primarily from metastatic site).
I am planning to flow breast cancer cells, in a buffer, into a device. What is the highest flow rate I could use if I want the breast cells to attract to the CXCL12 chemokine? Specifically, I am putting the CXCL12 on the side of the device and flowing the cancer cells in the middle. Will the force of attraction be strong enough to eventually have all the cancer cells on the side of the device with the chemokines?
I am using patients derived xenograft to study genes regulating breast cancer metastasis to the lung. So I will need to collect human breast cancer cells from the mouse lung.
Thus I will need a protocol for lung dissociation and selection for only human cancer cells.
I know of oncomine and TCGA. I am trying to see if a gene of my interest in involved in metastasis.
In various literature reports, CXCR4 interacting with CXCL12 has been said to cause metastasis in breast and prostate cancers. In colorectal cancers, reports have stated that the cancer cells respond to CCL20, meaning they contain the CCR6 receptor. However, shouldn't these three cancer cell lines all have the same receptors, since they are all epithelial in origin?
Circulating tumor cells (cancer cells that cause metastasis) respond to a variety of chemokines, causing it to travel through the bloodstream into specific organs. Which chemokines attract ONLY these cancer cells without having any effect on normal white blood cells?
Two big issues remaining for prostate cancer migration are cancer formation from hyperplasia and the way cancer to migrate. We may figure out if we can detect the beginning of different signaling of metastasis or lymphastasis.
Perineural invasion is frequently observed in pancreatic cancers and they exhibit focal differentiation surrounding the peripheral nerves. Neighboring schwann cells lead to the differentiation of invasive tumor cells and the formation of the tubule-like structures. The nerve growth factors are responsible for this peri-neural invasion with differentiation?
The co-culture experiment in combination of schwann cells with pancreatic cancer cells in vitro leads to the enhanced migration potential as well as higher levels of differentiation molecule expression in a heterogeneous way. It is noteworthy that the co-culture system without cell-to-cell attachment cannot induce the same result, indicating the possibility that neither soluble growth factors nor exosomes containing micro-RNA cannot be the cause of the alteration of tumor cell phenotype.
Although CTCs have a clinical impact in several tumor types like breast cancer or colorectal cancer, the role of CTCs in lung cancer remains unclear. However, I am planning a project in the area of CTCs in NSCLC and I'm on the search for the researchers who have experience with CTCs and are willing to participate. Feedback and opinions on the topic are also welcome.
I'm planning to use BrdU to label cancer cells in vitro, and inject the cell in vivo, in order to track the cell. For this, I want to know whether BrdU affects the biology of cancer cell or not..(especially metastasis)..I have done some searching on this, but I couldn't find any...
The only thing I found is a study which used cell tracker for this,[Mol Cancer. 2015 Dec;14(1):282] , These guys have done exactly what I am trying to do.
;however, as I know, fluorescent of cell tracker only lasts few weeks,,
That's why I am concerning BrdU
Q1. Would BrdU affect biological behaviour of cancer cells?
Q2. Does anybody know any study using BrdU-treated cancer cells for metastasis experiment?
Q3. Cell tracker vs. BrdU, for this experiment which one would you recommend ?
Can be Bone, Brain, Liver, Kidney, Peritoneal zone and Adrenal Glands? These six because have the propriety of "encapsulate" the tumors? I hope your answers, Thanks.
I want to know names of the most commonly and commercially available drugs against cancer metastasis, can anyone help?
I am going to perform a flow cytometry assay in order to detect myeloid cells and melanoma metastatic cells in the lung. For myeloid cells I am going to use CD11b antibody conjugated Alexa 488 and for melanoma cells Tyrosinase antibody conjugated with phycoerythrin. I am wondering what should be the positive and negative controls for gating in this assay.
KRAS,NRAS,BRAF wild type colorectal cancer can response the anti-EGFR therapy using cetuximab or panitumumab. However, in some cases such as Her2 or MET amplification there will be an resistant to anti EGFR therapy although tumor has KRAS,NRAS and BRAF wild type genotype. If there is a polysomy in chromosome 17(Her2:CEP17 ratio 1, however there is a increasing number of chromosome 17 such as 3:3,4:4), using FOLFIRI+Cetuximab+Herceptin can overcome EGFR resistant in metastatic colorectal cancer?
Prospective fate maps show that cells of different germ layers and different subdivisions within germ layers are partially segregated from one another in the epiblast and primitive streak. however , Gerhart et al (2007) demonstrate that the epiblast contains both multipotent cells and a subpopulation of cells that are stably committed to the skeletal muscle lineage before the onset of gastrulation .
Monte et al. suggest that pattern of metastatis in malignant melanoma may related to the embryological derivation of tissues involved.
could there is specific interaction between deifferent segregation in epiblast or they exposed to related factor that may explain organ - specific metastases ?
this make sense or there is something i understood wrongly ?
Is the migration procedure initiated within the metastasized cells or is there a chemo/electrical signaling from the ECM/endothelial cells/blood stream which recruits cells into the blood stream?
Of course cells should have the potential to migrate, but what it makes cells posses a response like migration?
In migration/cell invasion assays, I've seen that a "chemoattractant" is used. This means that metastatic cell lines, on their own, can't migrate. They need a cue to start moving towards the blood stream.
Any insight on the topic is much appreciated.
there are studies show reciprocal interaction between cancer cell and stroma mediated by extracellular vesicles ( cancer cell release microvesicles affect stromal cells which in turn release microvesicles affect cancer cell) at primary tumor enviroment, in addition cancer cell release extracellular vesicles play a role in pre-metastatic niche formation .
but is there reciprocal interaction between primary tumor and pre-metastaticniche mediated by these vesicles ( i mean do cells in premetastatic niche also release extracellular vesicles affect primary tumor ?)
We are using arteries from humans for myographical studies, but we have problems during contraction when we use phenylphrine (PE)? But, the arteries contract normally when we use KCl.
In their article and communications with the editor [1, 2] S. Halabi and coauthors reported on predictive significance of pain interference scores and pain intensity for overall survival in patients with castration-refractory prostate cancer (CRPC). Overall 317 men with CRPC included in the study, and the median survival times for patients with pain intensity score of 5 or greater was significantly shorter than in men whose pain intensity score was less than 5 (8.76 months, 95% CI 6.88-11.30 months vs. 18.15 months 95% CI 15.47-20.20months, P<0.001) by univariate analysis. After adjusting for other prognostic factors, pain intensity remained a statistically significant predictor of OS with adjusted hazard ratio of death 1.61 (95% CI 1.21-2.13, P<0.001).
However, data in the literature on predictive role of pain intensity for survival in patients with bone metastases from solid tumours are scarce.
1. Halabi, S., et al., Pain predicts overall survival in men with metastatic castration-refractory prostate cancer. J Clin Oncol, 2008. 26(15): p. 2544-9.
2. Klepstad, P. and S. Kaasa, Predication of cancer patients' survival by pain interference items is not the same as evidence that pain intensity is a survival predictor. J Clin Oncol, 2008. 26(25): p. 4215-6; author reply 4216-7.
Does anyone have experience regarding the difference in metastatic potential of B16-F10 and B16-F1 cells? How much longer would B16-F1 cells need to cause a substantial number of metastasis in the mouse lung when injected i.v.?
I found quite different statements in literature. Some showing almost no metastasis, some showing a huge number of metastasis after 2 weeks.
Thanks in advance!
I am willing to do in vivo study (Xenograft) using 4T1 cell line (murine breast cancer cell line) and as I understand this cell line can highly metastasize to different organs. My goal of the study is to inhibit growth and not stop metastasis. So, even though the cell line grows very fast, it is not the suitable. Any help will be appreciated.