Effect and mechanism of TMS on proliferation and apoptosis in human endometrial carcinoma cell line Ishikawa

Objective To investigate the effect and mechanism of 2,3′,4,5′-tetramethoxystilbene(TMS),the inhibitor of Cytochrome P4501B1(CYP1B1),on the proliferation and apoptosis in human endometrial carcinoma cell line Ishikawa.Methods Immunocytochemistry assay was applied to detect the expression of CYP1B1 protein.MTT method was used to measure cell proliferation after Ishikawa cells were treated with TMS,and the morphology of the cells was observed by inverted microscope.Flow cytometry was employed to investigate cell apoptosis,cell cycle and the expression of Bcl-2,Bax and Survivin.Results The expression of CYP1B1 protein was found in Ishikawa cells.TMS could inhibit the proliferation of Ishikawa cells significantly in a dose and time dependent manner(P0.05),and degeneration and necrosis of Ishikawa cells were found under inverted microscope.At the same time,the typical morphologic changes of cell apoptosis appeared,such as cell nucleus concentration,nuclear fragmentation and so on.TMS could promote the apoptosis of Ishikawa cell,and the ratio of cell apoptosis was increased with the concentration of TMS.Cell cycle phase analysis revealed that the proportion of Ishikawa cells in G0/G1 phase decreased,while the proportion of Ishikawa cells in G2/M phase increased in a concentration-dependent manner(P0.05).The expression of Bcl-2 and Survivin was down-regulated and the expression of Bax was up-regulated in a concentration-dependent manner(P0.05).Conclusion TMS can inhibit the proliferation of human endometrial carcinoma Ishikawa cells,induce its apoptosis and block the Ishikawa cell at G2/M phase.The mechanism may be through down-regulation of Bcl-2 and Survivin,and the up-regulation of Bax.