Department of Gynecology, Harbin Medical University Cancer Hospital, Harbin, China.
Department of Surgery, Harbin Medical University Cancer Hospital, Harbin, China.
Exp Cell Res. 2020 Nov 1;396(1):112241. doi: 10.1016/j.yexcr.2020.112241. Epub 2020 Aug 21.
Epithelial-mesenchymal transition (EMT) is an important contributor to drug resistance in ovarian cancer. The aims of this study were to explore the potential role of the miR-302 cluster in modulating EMT and cisplatin resistance in ovarian cancer. We used qRT-PCR and western blotting to show that miR-302 expression was lower in chemoresistant than in chemosensitive cells, and miR-302 was upregulated in chemosensitive, but not chemoresistant ovarian cancer cells in response to cisplatin treatment. We identified ATAD2 as a target of miR-302 and showed that ectopic expression of miR-302 increased cisplatin sensitivity and inhibited EMT and the invasiveness of cisplatin-resistant cells in vitro by targeting ATAD2. Knockdown of ATAD2 restored cisplatin sensitivity and reversed EMT/metastasis in cisplatin-resistant cells, as shown by western blotting and invasion/migration assays. The effect of miR-302 overexpression on EMT and invasiveness was mediated by the modulation of β-catenin nuclear expression. Immunofluorescence analysis showed that ATAD2 overexpression reversed the miR-302-induced downregulation of nuclear β-catenin in cisplatin resistant cells. A xenograft tumor model was used to show that miR-302 increases the antitumor effect of cisplatin in vivo. Taken together, these results identify a potential regulatory axis involving miR-302 and ATAD2 with a role in chemoresistance, indicating that activation of miR-302 or inactivation of ATAD2 could serve as a novel approach to reverse cisplatin resistance in ovarian cancer.
上皮-间充质转化(EMT)是卵巢癌耐药的重要原因。本研究旨在探讨 miR-302 簇在调节卵巢癌 EMT 和顺铂耐药中的潜在作用。我们使用 qRT-PCR 和 Western blot 表明,耐药细胞中 miR-302 的表达低于敏感细胞,顺铂处理后敏感细胞而非耐药细胞中 miR-302 上调。我们确定 ATAD2 是 miR-302 的靶标,并表明外源性表达 miR-302 通过靶向 ATAD2 增加顺铂敏感性并抑制 EMT 和耐药细胞的侵袭性。ATAD2 的敲低恢复了顺铂耐药细胞的顺铂敏感性并逆转了 EMT/转移,Western blot 和侵袭/迁移实验证实了这一点。miR-302 过表达对 EMT 和侵袭性的影响是通过调节β-连环蛋白核表达介导的。免疫荧光分析表明,ATAD2 过表达逆转了 miR-302 诱导的耐药细胞中核β-连环蛋白的下调。异种移植肿瘤模型表明 miR-302 增加了顺铂在体内的抗肿瘤作用。总之,这些结果确定了涉及 miR-302 和 ATAD2 的潜在调节轴在化疗耐药中的作用,表明激活 miR-302 或失活 ATAD2 可能成为逆转卵巢癌顺铂耐药的新方法。