异常甲基化的原钙黏蛋白7对雄激素非依赖性前列腺癌细胞生长的影响

Effect of abnormal methylation of PCDH7 on growth of androgen-independent prostate cancer cells

目的探讨异常甲基化的原钙黏蛋白7(PCDH7)对雄激素非依赖性前列腺癌(AIPC)细胞生长的影响。方法培养AIPC细胞系LNCaP-AI,采用甲基化酶抑制剂5-氮杂-2′-脱氧胞苷(AZA)处理细胞24 h,RT-PCR和Western blot法检测PCDH7 mRNA和蛋白的表达水平,CCK-8法检测细胞生长情况,流式细胞术检测细胞凋亡,Transwell法检测细胞侵袭能力。结果加入AZA后,PCDH7基因启动子片段1甲基化水平为0.00486±0.00144,低于对照组的0.00956±0.00594,片段2甲基化水平为0.00610±0.00171,低于对照组的0.00858±0.00180(P<0.05),且PCDH7 mRNA和蛋白表达均上调(P<0.01)。加入DNA甲基化转移酶1后PCDH7 mRNA和蛋白表达下调(P<0.05)。加入AZA后,细胞生长抑制率和细胞凋亡率增加[(15.3%vs.0%)和(5.06%vs.3.44%)](P<0.01),侵袭细胞数下降(P<0.01)。结论异常甲基化的PCDH7在AIPC细胞中表达上调,能够抑制细胞生长和侵袭能力,促进细胞凋亡。

Objective To investigate the effect of abnormal methylation of procalcitonin 7(PCDH7)on the growth of androgen independent prostate cancer(AIPC)cells.Methods AIPC cell line LNCaP-AI was cultured and treated with 5-aza-2′-deoxycytidine(AZA)for 24 hours.RT-PCR and Western blot were used to detect the mRNA and protein expressions of PCDH7.The cell growth was detected by CCK-8 assay.The cell apoptosis was detected by flow cytometry.The cell invasion was detected by Transwell assay.Results The methylation level of PCDH7 promoter fragment 1 was 0.00486±0.00144 after the treatment of AZA,which was lower than 0.00956±0.00594 of the control(P<0.05).The methylation level of PCDH7 promoter fragment 2 was 0.00610±0.00171 after the treatment of AZA,which was lower than 0.00858±0.00180 of the control(P<0.05).The mRNA and protein expressions of PCDH7 were increased after the treatment of AZA(P<0.01),while those were decreased after the treatment of DNA methyltransferase 1(P<0.05).The inhibition rate of cell growth and the rate of cell apoptosis were increased(15.3%vs.0%and 5.06%vs.3.44%)(P<0.01),and the number of invasion cells was decreased(P<0.01).Conclusion The abnormal methylation of PCDH7 is increased in AIPC cells,which could inhibit the cell growth and invasion and promote the cell apoptosis.