Experimental Study Of Phenylethyl Isothiocyanate On Bladder Cancer Cells Proliferation And OPCML Expression

Objective To study the effect of phenylethyl isothiocyanate(PEITC)on the expression of bladder cancer suppressor gene OPCML,and to explore its effect on cell proliferation.Methods The bladder cancer line T24 was cultured in vitro,and incubated with different dose of PEITC.MTT was used to observe the effect of PEITC on the cell proliferation,RT-PCR and Western blot were used to analyze the mRNA and protein expression level,respectively.Methylation-specific PCR was used to detect the methylated level of the promoter district of OPCML,qT-PCR and Western blot were used to detect DNMT1 mRNA and protein expression.Xenografts in nude mice was used identify the effect of PEITC on xenografts growth,and the expression of OPCML and DNMT1 were analyzed.Results: 1.MTT results demonstrated that 1,10,30 and 50μmol/L of PEITC could inhibit the proliferation in dose-dependent manner;2.qPCR results showed the mRNA of OPCML in the control group was very low,while increased dramatically after incubated with 1,10,30 and 50μmol/L of PEITC.Similar results were obtained by Western blot;3.MSP results showed PEITC could inhibit the methylation of OPCML in the promoter district,and the methylatedactivity in the T24 cells.In addition,PEITC treatment could decrease the expression of DNMT1 mRNA and the protein level;4.PEITC could inhibit the growth of the Xenografts in nude mice,down-regulation of the expression of DNMT1 and up-regulation of the OPCML expression in the Xenografts.Conclusion: 1.PEITC could suppress the proliferation of T24 cell and the growth of xenografted tumor;2.PEITC upregulate the expression the OPCML by suppressing the methylated activity in T24 cells...