姜黄素增强PTEN野生型三阴性乳腺癌细胞对PARP1抑制剂敏感性的研究

Curcumin sensitizes wild-type PTEN breast cancer cells to PARP1 inhibitors

目的研究姜黄素增强PTEN(磷酸酶和张力蛋白同系物)野生型三阴性乳腺癌细胞对于PARP1抑制剂奥拉帕尼(Olaparib)的敏感性及其潜在的作用机制,从而为PTEN野生型三阴性乳腺癌病人的临床治疗提供新方案。方法使用PTEN缺陷三阴性乳腺癌细胞株BT549和PTEN野生型细胞株BT549-PTEN进行实验。MTT实验检测两种细胞在奥拉帕尼作用下的存活率。蛋白质印迹实验检测PTEN缺陷型和野生型细胞中同源重组修复关键蛋白RAD51的表达。彗星实验检测姜黄素及姜黄素与奥拉帕尼联合使用后细胞DNA的损伤程度。蛋白质印迹检测姜黄素作用后RAD51变化。结果PTEN可通过增加RAD51的方式促进DNA损伤的修复。与PTEN缺陷型细胞相比,野生型细胞对于奥拉帕尼的作用较不敏感。姜黄素能够通过抑制RAD51的方式抑制PTEN介导的DNA修复,并增强PTEN野生型三阴性乳腺癌对奥拉帕尼的敏感性。结论姜黄素通过抑制PTEN介导的DNA修复使PTEN野生型乳腺癌细胞对PARP1抑制剂奥拉帕尼敏感。

Objective This research aimed to:(1)investigate the effects of curcumin in phosphatase and tensin homolog(PTEN)-mediated DNA repair and in the sensitization of wild-type PTEN triple-negative breast cancer cells to Olaparib,a poly ADP ribose polymerase(PARP)1 inhibitor;and(2)provide a new therapeutic strategy for patients with wild-type PTEN triple-negative breast cancer in the clinical setting.Methods Wild-type PTEN and PTEN-deficient variants of the triple-negative breast cancer cell line BT549 were used.MTT was used to assay the cell survival rate.Western blot was performed to detect the expression of RAD51(a key protein in homologous recombination repair)in wildtype PTEN cells.The comet assay was used to measure the DNA damage caused by curcumin and olaparib.Western blot was used to detect RAD51 after curcumin treatment.Results Wild-type PTEN cells were less sensitive to olaparib than PTEN-deficient BT549 cells.PTEN promoted DNA repair by increasing the expression of RAD51.Curcumin inhibited PTEN-mediated DNA repair by inhibiting RAD51 and enhanced the sensitivity of wild-type PTEN triple-negative breast cancer cells to olaparib.Conclusions Curcumin inhibited PTEN-mediated DNA repair and sensitized wild-type PTEN breast cancer cells to olaparib.