Objective To investigate the role and molecular mechanism of suvivin, an anti apoptosis gene, in cell growth and transformation. Methods Coding sequence of survuvin was amplified from Daudi cell mRNA by RT PCR and then cloned into prokaryotic and eukaryotic vectors. The vectors with survivin were transfected into BL21 cells of Escherichia coli and human embryonic kidney 293 cells. The cells were cultured. Protein was extracted from the cells and examined by gel electrophoresis. Suspension of 293 cells was cultured and the number of cells was counted every other day, thus a growth curve was drawn. Another suspension of 293 cells was cultured in soft agar to observe the number of colonies. Cells transfected with plasmids void of survivin were used as controls. Results The anti apoptosis gene survivin was well expressed in BL21 cells and 293 cells. The growth curve showed that the proliferation rate of 293 cells was slightly faster than that of control cells, hoever, without significant difference. Soft agar assay showed that the colonies formed by survivin transfected 293 cells were of greater size and with greater number. Western blotting showed overexpreaaion of cyclin D1 and c myc, two important cancer proteins, in cells transfected with survivin. Conclusion The anti apoptosis gene survivin promotes cell transformation. These effects may depend on the functions of cyclin D1 and c myc.
National Medical Journal of China
国家杰出青年基金资助项目( 3992 50 2 0 )
科学基金资助项目( 7992 0 35)