Sulforaphane enhances the cisplatin sensitivity through regulating DNA repair and accumulation of intracellular cisplatin in ovarian cancer cells
Sulforaphane enhances the cisplatin sensitivity through regulating DNA repair and accumulation of intracellular cisplatin in ovarian cancer cells
复制标题
DOI：
10.1016/j.yexcr.2020.112061
复制
发表时间：
2020-08-15
影响因子：
3.7
通讯作者：
Wu, Qi-Jun
中科院分区：
文献类型：
--
作者：
Gong, Ting-Ting;Liu, Xiao-Dong;Wu, Qi-Jun
目的：顺铂是一种广泛应用于包括卵巢癌在内的多种肿瘤的抗癌药物。但顺铂耐药的发生严重阻碍了顺铂的抗肿瘤作用。本研究旨在探讨莱菔硫烷（sulforaphane，SFN）对卵巢癌顺铂耐药的逆转作用及其机制。CCK-8法检测细胞活力。彗星试验检测DNA修复情况。用蛋白质印迹法检测顺铂转运蛋白的表达。HPLC法检测细胞内顺铂浓度。应用荧光素酶活性测定来确定miR-30 a-3 p在ERCC 1和ATP 7A的3 'UTR上的靶位点。结果：SFN通过诱导细胞DNA损伤和细胞内顺铂蓄积，逆转A2780/CP 70和IGROV 1-R10卵巢癌细胞对顺铂的耐药性。SFN处理显著增加miR-30 a-3 p表达，而顺铂耐药细胞中miR-30 a-3 p表达降低。miR-30 a-3 p过表达可增强A2780/CP 70和IGROV 1-R10细胞对顺铂的敏感性，抑制miR-30 a-3 p活性可减弱SFN对顺铂耐药的逆转作用。结论：SFN可通过上调miR-30 a-3 p的表达，诱导细胞DNA损伤，增加细胞内顺铂的蓄积，从而增强卵巢癌细胞对顺铂的敏感性。
Objectives: Cisplatin is commonly applied as anticancer agent for various cancers, including ovarian cancer. Unfortunately, the drug resistance frequently occurred which obstructing the effect of cisplatin on tumors. The goal of our research was to investigate the reversal actions and the potential mechanisms of sulforaphane (SFN) on cisplatin resistance in ovarian carcinoma.Methods: The A2780 and IGROV1 cells and their cisplatin resistance cells A2780/CP70 and IGROV1-R10 were used in this study. Cell viability was detected by CCK-8. The DNA repair was measured by comet assay. The cisplatin transporter proteins were measured with western blotting. The concentration of intracellular cisplatin was detected by HPLC. The luciferase activity assay was applied to determine the target site of miR-30a-3p on the 3'UTR of ERCC1 and ATP7A. A2780/CP70 and IGROV1-R10 xenograft mouse model were established to confirm the antineoplastic action of SFN combined with cisplatin.Results: SFN reversed the resistance of A2780/CP70 and IGROV1-R10 ovarian carcinoma cells to cisplatin through inducing DNA damage and accumulation of intracellular cisplatin. SFN treatment notably increased miR-30a-3p expression, which was decreased in cisplatin-resistant cells. Moreover, overexpressed miR-30a-3p enhanced the sensitivity of A2780/CP70 and IGROV1-R10 cells to cisplatin treatment, and inhibiting miR-30a-3p activity abated the reversal actions of SFN on cisplatin resistance. The luciferase assay findings showed that miR-30a-3p binds to ERCC1 and ATP7A which are the key regulators for DNA repair and cisplatin transportation.Conclusions: Our findings indicated that SFN could enhance cisplatin sensitivity of ovarian carcinoma cells through up-regulating miR-30a-3p to induce DNA damage and accumulation of intracellular cisplatin.