地西他滨抑制前列腺癌细胞的增殖和迁移侵袭

The effect of decitabine on the biological behavior of prostate cancer cells

目的:探究DNA甲基转移酶1(DNMT1)抑制剂地西他滨(DAC)对前列腺癌PC3细胞的增殖和迁移侵袭的影响。方法:取对数生长前列腺癌细胞株PC3,分为3组,分别为对照组和实验组(DAC处理组,250 nmol/L和500 nmol/L)。采用细胞计数试剂盒(CCK-8)和单克隆检测细胞体外增殖能力;Transwell检测PC3细胞体外迁移和侵袭能力的改变。定量聚合酶链反应(qPCR)和蛋白质印迹法(Western blot)检测DAC对DNMT1的影响效率,并检测DAC处理后p21水平改变。组间比较采用t检验。结果:DAC使用后显著抑制PC3细胞增殖能力,并呈剂量依赖性。CCK-8实验显示Ctrl组、250 nmol/L DAC和500 nmol/L DAC组的吸光度值分别为1.831±0.105、1.530±0.119和1.329±0.056,与对照组比较实验组差异均有统计学意义(t=3.247,P<0.05;t=7.299,P<0.05)。单克隆实验结果显示Ctrl组、250 nmol/L DAC和500 nmol/L DAC克隆形成数目为(827.67±4.04)、(577.33±2.52)和(283.67±6.03)个,与对照组比较实验组差异均有统计学意义(t=91.072、129.835,P<0.05)。Transwell结果显示,DAC处理组细胞迁移和侵袭能力低于对照组(P<0.05)。Western blot和qPCR结果显示,DNMT1表达随DAC剂量增高表达而逐渐下降,呈剂量依赖性。DAC处理后,PC3细胞的p21表达水平呈剂量依赖性增高。结论:DAC能够有效抑制前列腺癌PC3细胞的增殖、迁移和侵袭能力,通过调节DNMT1-p21信号通路可能是作用机制之一。

Objective To explore the effect of DNA methyltransferase 1(DNMT1)inhibitor decitabine(DAC)on the proliferation,migration and invasion of prostate cancer PC3 cells,and explore its mechanism.Methods The logarithmic prostate cancer cell line PC3 was divided into three groups:control group and experimental group(DAC treatment group,250 nmol/L and 500 nmol/L),and the in vitro proliferation ability of the cells was detected by cell counting kit-8(CCK-8)and monoclonal assay;Transwell assay was used to detected the changes of different doses of DAC on the migration and invasion ability of PC3 cells.The effect of DAC on DNMT1 and p21 was detected by quantitative polymerase chain reaction(qPCR)and Western blotting.T-test was used for comparison between groups.Results DAC significantly inhibited the proliferation of PC3 cells in a dose-dependent manner.The CCK-8 experiment showed that the absorbance of the Ctrl group,250 nmol/L DAC and 500 nmol/L DAC groups were 1.831±0.105,1.530±0.119 and 1.329±0.056,respectively,and the difference between the experimental group and the control group was statistically significant(t=3.247,P<0.05;t=7.299,P<0.05).The monoclonal experiment showed that the Ctrl group,250 nmol/L DAC and 500 nmol/L DAC clone formation numbers were 827.67±4.04,577.33±2.52 and 283.67±6.03,and the difference between the experimental group and the control group was statistically significant(t=91.072,129.835,P<0.05).The results of Transwell experiments showed that,the ability of migration and invasion of cells in the DAC treatment group was reduced compared with Ctrl group.The results of Western blotting and qPCR showed that the expression of DNMT1 decreased and p21 increased with the treatement with DAC.Conclusion DAC can effectively inhibit the proliferation,migration and invasion of prostate cancer PC3 cells.The mechanism of action may be through the regulation of DNMT1-p21 signaling pathway experiments.