I want to study various signaling pathways that are involved in cancer.
Dear Kiran
Yes, somatic cell lines can be transformed using "transforming viruses" [such as for example, certain strains of adenoviruses,] radiation and chemical carcinogens. Please review studies done in this area during 60s and 70s
It depends...
Do you want to transform rodent or human cells. Mouse cells can be easily transformed. Human cells are very resistant to transformation in vitro.
To my knowledge, the only reliable recipe was published by the Weinberg group. However, it later turned out that they mistakenly introduced the whole "early region" of SV40, instead of just large-T, as claimed in in Nature. (By the way, Nature never published a correction.)
1. Hahn WC, Counter CM, Lundberg AS, Beijersbergen RL, Brooks MW, Weinberg RA. Creation of human tumour cells with defined genetic elements. Nature 1999; 400(6743): 464-8.
It'sby now well known that many different stimuli, such as chemical, physical, infectious (in particular viruses) may be responsible of somatic cell transformation.
Read the Weinberg paper, it says it all. Cell transformation of normal HUMAN cells (with a diploid karyotype) is a very inefficient process and only rarely works. Thats the reason why everybody is using human cancer cell lines, instead of producing their own human cancer cells in vitro...
The best example of oncogenic transformation with high efficiency of somatic cells is the EBV transformation of B cells with the mean of EBV virus. There are also other examples similar in both human and mouse as mentioned above.
Please keep in mind that in a natural occurring cancer many conditions are required in order to allow cancer cells to thrive and resist to the immune system. Therefore, it is very likely that a pure in vitro system will never resemble to the real cancer you may find in a patient.
EBV transformed B cells are called lymphoblastoid cell lines, and they are immortal...but not completely transformed. After many generations in cell culture they eventually transform...
Yes, as above, many viruses such as EBV or Sendai virus may be useful in vitro. Methylcholanthrene can induce fibrosarcomas in mice which can then be grown in culture.
I want to induce cancer by Chemical carcinogen 7,12-dimethylbenz[a]anthracene (DMBA).
If you do, I am afraid you will use human fibroblasts. However, cancer is usually a disease of epithelial tissues. Carcinomas account for about 80% of cancer deaths.
So if you study fibroblasts, they won't tell you much about carcinomas. On the other hand, if you want to transform primary human epithelial cells with DMBA, I wish you luck...you will need it ;-)
Thank you so much for your nice discussion, As you said, Human cells are very resistant to transformation in vitro, How about Hamster somatic cell lines R1610?. I have seen few papers on R1610 . No one has done done with chemical carcinogen with these cell lines.
http://www.ncbi.nlm.nih.gov/pubmed/8694278
http://www.ncbi.nlm.nih.gov/pubmed/3047139
You may try using tumor cells from syngeneic tumors induced by chemical carcinogens and study them. There are also tumor cell lines available in various cell bank,alsom commercially available.You may like to procure some of these cell lines from them and study the signalling pathways
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