Capture and characterization of 5-aza-2'-deoxycytidine-treated C3H/10T1/2 cells prior to transformation

摘要: Abstract A model system was developed to capture phenotypically normal cells committed transformation address two fundamental questions in cancer biology: (i) what are the earliest events transformation; and (ii) is role of DNA methylation carcinogenesis? Individual C3H/10T1/2 were treated with 5-aza-2'-deoxycytidine, which causes hypomethylation DNA. Cells grown subconfluence, individual microcolonies trypsinized into fractions. One fraction cryopreserved, other replated maintained culture. Ten percent these colonies became morphologically transformed after 4-6 weeks. The cryopreserved ancestral both nontransformed then cultured compared each transformants for phenotypic properties cellular transformation. Pretransformed nonpretransformed initially indistinguishable, their early growth curves did not differ significantly, they form soft agar. On continued culture, however, pretransformants displayed all characteristics Furthermore, occurred a given pretransformant most or that clone. Thus, could be isolated this way prior Studies pretransformed will permit examination carcinogenesis