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山东大学学报(医学版) ›› 2016, Vol. 54 ›› Issue (5): 6-11.doi: 10.6040/j.issn.1671-7554.0.2015.648

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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

刘相永,孙颖,黎莉   

  1. 山东大学齐鲁医院(青岛)放疗科, 山东 青岛 266035
  • 收稿日期:2015-07-03 出版日期:2016-05-16 发布日期:2016-05-16
  • 通讯作者: 黎莉. E-mail:drlili5060@163.com E-mail:drlili5060@163.com

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

LIU Xiangyong, SUN Ying, LI Li   

  1. Department of Radiotherapy, Qilu Hospital of Shandong University, Qingdao 266035, Shandong, China
  • Received:2015-07-03 Online:2016-05-16 Published:2016-05-16

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。

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

Abstract: 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 line 22RV1 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 TM2000A, respectively. The viability of cells treated with PARP inhibitor was measured by CCK-8 assay. Cell distribution of cell cycle was determined by flow cytometry. Levels of BRCA1, PTEN, RAD51, PARP, and NF-κB protein were measured by Western blotting. Results PARP inhibitor AG014699 inhibited the proliferation of MCF-7 with 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 山 东 大 学 学 报 (医 学 版)54卷5期 - 刘相永,等.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 \=-could markedly inhibit the proliferation of cells possibly due to the attenuation of RAD51 and NF-κB activity.

Key words: Poly(ADP-ribose)polymerase, Breast cancer susceptibility genes, Nuclear factor κB, RAD51, Phosphatase and tensin homology deleted on chromosome ten

中图分类号: 

  • R73-36+1
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