PARP-1抑制剂对BRCA1基因沉默的乳腺癌细胞株MCF-7及PTEN基因沉默的前列腺癌细胞株22RV1增殖的影响  

The effects of poly(ADP-ribose) polymerase inhibitor on the proliferation of breast cancer cell line MCF-7 and prostate cancer cell line 22RV1 with BRCA and PTEN gene silencing

作　　者：刘相永 孙颖[1] 黎莉[1] 

机构地区：[1]山东大学齐鲁医院(青岛)放疗科,山东青岛266035

出　　处：《山东大学学报（医学版）》2016年第5期6-11,共6页Journal of Shandong University:Health Sciences

摘　　要：目的探讨多聚腺苷二磷酸核糖聚合酶-1(PARP-1)抑制剂对被沉默人类乳腺癌易感基因(BRCA1)的乳腺癌细胞株MCF-7及被沉默张力蛋白同源10号染色体丢失的磷酸酶基因(PTEN)的前列腺癌细胞株22RV1的作用。方法利用慢病毒介导的RNA干扰技术构建人乳腺癌MCF-7细胞的BRCA-1基因沉默细胞株及前列腺癌22RV1细胞的PTEN基因沉默细胞株。CCK-8法、流式细胞术分别检测PARP抑制剂AG014699对MCF-7、22RV1及转染细胞的增殖及周期分布的影响。Western blotting印迹法检测BRCA1、PTEN、DNA损伤修复酶RAD51、PARP、核因子κB(NF-κB)的蛋白表达。结果被沉默BRCA1基因的MCF-7细胞株及被沉默PTEN基因的22RV1细胞不表达RAD51。PARP抑制剂可明显抑制BRCA1基因沉默的MCF-7细胞及PTEN基因沉默的22RV1细胞的增殖,呈浓度时间依赖性,可能与RAD51、NF-κB表达降低有关;PARP抑制剂使细胞周期阻滞于G2/M期。结论 PARP抑制剂能抑制存在DNA同源重组修复缺陷细胞(由BRCA1及PTEN功能异常所导致)的增殖,该作用可能与RAD51、NF-κB表达降低有关。Objective To evaluate the proliferative effects of poly（ ADP-ribose） polymerase-1（ PARP-1） inhibitor on breast cancer cell line MCF-7 with breast cancer susceptibility（ BRCA） genes silencing and prostate cancer cell line22RV1 with phosphatase and tensin homology deleted on chromosome ten（ PTEN） silencing. Methods BRCA1 /PTEN-targeted small interfering double-stranded RNAs（ siRNA） were introduced into MCF-7 /22RV1 cells by Lipofectamine TM2000 A,respectively. The viability of cells treated with PARP inhibitor was measured by CCK-8 assay.Cell distribution of cell cycle was determined by flowcytometry. Levels of BRCA1,PTEN,RAD51,PARP,and NF-κB protein were measured by Western blotting. Results PARP inhibitor AG014699 inhibited the proliferation of MCF-7with BRCA1 genes silencing and 22RV1 with PTEN silencing in a concentration and time-dependent manner,and blocked the cell cycle in G2 / M. RAD51 and NF-κB protein expressions in the proliferation of MCF-7 with BRCA1 genes silencing and 22RV1 with PTEN silencing were decreased. Conclusion Significant growth inhibition could be observed upon PARP inhibitor AG014699 in cells with BRCA1 and PTEN-associated DNA repair defects,indicating that AG014699 could markedly inhibit the proliferation of cells possibly due to the attenuation of RAD51 and NF-κB activity.

关 键 词：多聚腺苷二磷酸核糖聚合酶 人类乳腺癌易感基因 张力蛋白同源10号染色体丢失的磷酸酶基因 RAD51 NF-κB信号通路 

分 类 号：R155.3[医药卫生—营养与食品卫生学]