I want to transform normal cell into cancerous ones in cell culture. We do not have animal facility to check tumor formation in mice/ rats. so I have Four questions:
1) How can i convert normal cells into cancerous ones in cell culture? Anyone has experience treating normal cells with Benzo(a)Pyrene/ Aflatoxin B1 like carcinogens and convert cells into cancerous ones?
2) Are these Chronic treatments (low dose treatments for weeks) or just one time treatments?
3) In such experiments how do you detect that cells have become cancerous? Does they form colonies/ foci? Does Soft Agar Colony Formation assay do justice here?
4) What % of normal cells can be transformed into cancerous this way?
It would be helpful if you would indicate whether you would like to "transform" human cells or rodent cells. Because the steps that are involved are quite different and could be challenging.
@Divaker Choubey, i'm using normal human cells such as WI38, MCF12F etc
WI-38 human lung fibroblasts have been immortalized using SV40 virus. The virus targets both p53 and Rb tumor suppressors. However, these two events are not sufficient to acquire the transformed phenotype. Therefore, activated Ras oncogene may be needed to make cells serum-independent, contact inhibition independent, and form colonies in soft agar (all associated with a transformed phenotype). Formation of colonies in soft agar is a requirement for forming a tumor in animals.
If anyone is able to "simply" transform a normal cell line into a cancerous one, for sure she / he will rapidly earn a Nobel Prize ...
Cancer biology is "little bit" more complicated ... and you should be by far much more precise about your "wishes" in transforming "normal cells into cancer ones" ...
The attached articles (if you are reading them "in depth") will may be indicate you that your wish(s) is/are not very simple ...
Best regards
Robert
PS: I forgot to ask you whether you want a "very aggressive cancer cell model prisoneer in a plastic flask" ... or if you want "more or less mimicing the cilinical situation" ... because unfortunately for cancer patients, their cancer does not develop in a plastic flask but highly benefit from thir tumor microenvironment ...
I am sure that Divaker is perfectly aware of all these points.
Retroviral transduction of SV40 T antigen, hTERT, and activated H-ras into WI-38 cells has been shown to lead to anchorage independence in soft agar.
https://www.ncbi.nlm.nih.gov/pubmed/14563552
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