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目的研究三氧化二砷(arsenic trioxide,As2O3)对激素非依赖性前列腺癌DU145细胞的生长抑制作用及对RASSF1A基因去甲基化和蛋白表达的影响。方法应用MTT法检测不同浓度(0.5、1.0、2.0、4.0、6.0、12.0、20.0μmol/L)的As2O3不同作用时间(24、48、72 h)对DU145细胞的生长抑制作用;应用甲基化特异性PCR(MSP)和West-ern blot检测As2O3对DU145细胞RASSF1A基因甲基化状态及蛋白表达的影响。结果 As2 O3可抑制DU145细胞的增殖,在一定范围内随着药物浓度的增高,抑制作用逐渐增强(F=838.089,P<0.05);同一浓度作用时间越长,抑制率越高(F=8.849,P<0.05);且As2O3可使RASSF1A基因甲基化逆转,蛋白重新表达。结论 As2O3可以逆转前列腺癌DU145细胞RASSF1A基因启动子CpG岛的异常甲基化,诱导该抑癌基因的重新表达,抑制前列腺癌DU145细胞的增殖。
Objective To study the effects of arsenic trioxide (As2O3) on the growth of hormone-independent prostate cancer DU145 cells and its effect on the demethylation and protein expression of RASSF1A gene. Methods The growth inhibition of DU145 cells treated with different concentrations of As2O3 (0.5, 1.0, 2.0, 4.0, 6.0, 12.0 and 20.0 μmol / L) for 24 h, 48 h and 72 h was detected by MTT assay. The effect of As2O3 on the methylation status and protein expression of RASSF1A gene in DU145 cells was detected by MSP and West-ern blot. Results As2 O3 inhibited the proliferation of DU145 cells. The inhibitory effect was gradually enhanced with the increase of drug concentration in a certain range (F = 838.089, P <0.05). The longer the same concentration was, the higher the inhibitory rate was (F = 8.849 , P <0.05). As2O3 could reverse the methylation of RASSF1A gene and re-express the protein. Conclusion As2O3 can reverse abnormal methylation of RASSF1A gene promoter CpG island in prostate cancer DU145 cells, induce the re-expression of this tumor suppressor gene, and inhibit the proliferation of prostate cancer DU145 cells.