A eukaryotic vector highly expressing PKCγ,was constructed by DNA recombination and transfected into C3H10T1/2 cells by gene transfection. The NCP4 cells stably expressing PKCγ was isolated successfully by Southern blot,Western blot and PKC activity assay.The NCP4 cells displayed an enhanced growth rate,especially under low serum conditions, apparently reduced dependence on serum. By using FCM, it was shown that NCP4 cells exhibited decreased percentage in G1 phase, increased percentage in S phase and G2+M phase. In contrast to control cells, NCP4 cells overexpressing PKCγ decreased anchorage dependence and formed small colonies in soft agar. Furthermore, it was observed that the expression of oncogene c-sis increased obviously in NCP4 cells, it may be one of the molecular mechanisms of the reduced dependence on serum in NCP4 cells. The results indicated that specific elevation of the PKCγ level directly affected the increase of growth rate and lead to some transforming phenotypes in C3H10T1/2 cells.
ZHAO Xia, LIU Hui-Tu. Preliminary Analysis of Disordered Growth Control in C3H10T1/2 Cells Caused by Overexpression of PKCγ[J]. Progress in Biochemistry and Biophysics,1999,26(6):569-573
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