摘要
Expression of DNA methyltransferase 1(DNMT1),which plays an important role on aberrantly methylated CpG in the promoter regions of tumor sup-pressor genes(TSGs),is higher in bladder cancer cells than in normal bladder cells.Therefore,its overexpression is closely related to tumor formation.In this study,the eukaryotic vector pshRNA-DNMT1 was constructed and transfected into T24 cells.Levels of DNMT1 mRNA and protein were detected by reverse transcription-polymerase chain reaction(RT-PCR)and western blot.Relative to the blank control at the 24th,48th and 72nd hour after transfection of pshRNA-DNMT1,the inhibitory rates of DNMT1 mRNA levels in T24 cells were 28.44%,52.48%,70.91%,respectively.Those of DNMT1 proteins were 24.27%,57.79%,and 77.74%,respectively.Proliferation and apoptosis were assayed by MTT and flow cytometry with Annexin-V-FITC/PI staining.The growth inhibition rates of pshRNA-DNMT1 at the 24th,48th and 72nd hour after transfection of pshRNA-DNMT1 were(4.34¡0.76)%,(9.87¡1.54)%and(13.78¡1.93)%,respectively.There were statistically significant differ-ences between pshRNA-DNMT1 and the control blank at each time points(P,0.01);24,48 and 72 hours after T24 cells were transfected by pshRNA-DNMT1,the apoptosis rates of pshRNA-DNMT1 were(3.87¡0.81)%,(8.69¡1.23)%and(11.46¡1.24)%,respectively(P,0.01 vs blank control).Based on this case,our conclu-sion is that the recombinant plasmid pshRNA-DNMT1 can silence the expression of gene DNMT1 mRNA and protein effectively,and to some extent,it also can inhibit the proliferation of bladder cancer cell and promote the cellular apoptosis.
