Tian Xueye, Zuo Xiaohang, Hou Meng, Li Chen, Teng Yue
Department of Obstetrics and Gynaecology, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, China.
Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
J Cancer. 2021 Jul 25;12(19):5712-5722. doi: 10.7150/jca.58979. eCollection 2021.
Platinum-based chemotherapy is part of current standard treatment for epithelial ovarian cancer (EOC). However, chemoresistance often rapidly developed, leading to chemotherapy failure and unfavored prognosis. Increasing evidence has demonstrated the important role of oncogenic long noncoding RNA H19 in various cancers, including EOC. No current study is available in exploring the role of lncRNA-H19 in carboplatin resistance of EOC and its underlying mechanism. Levels of lncRNA-H19, miR-29b-3p, and STAT3 mRNA were measured by qRT-PCR. The 50% inhibitory concentration value was detected with Cell Counting Kit-8 (CCK8). Colony-formation and CCK8 assays were employed to measure cell viability. Cell migration and invasion ability was evaluated with transwells. Western blot assay was utilized to measure P-gp, MRP1, LRP, and STAT3 protein levels. The targeting between lncRNA-H19 and miR-29b-3p, as well as miR-29b-3p and STAT3, was verified by dual-luciferase, RNA immunoprecipitation, and RNA pull-down experiments. lncRNA-H19 and STAT3 were sharply increased, while miR-29b-3p was decreased in carboplatin-resistant EOC. Carboplatin efficacy was enhanced by lncRNA-H19 silencing in chemo-resistant EOC cells. lncRNA-H19 served as a competing endogenous RNA of miR-29b-3p, causing the derepression of miR-29b-3p downstream target STAT3, leading to chemoresistance in carboplatin-tolerated EOC. The lncRNA-H19/miR-29b-3p axis improved carboplatin resistance of EOC by targeting STAT3, indicating a possible approach to improving chemotherapy for EOC.
铂类化疗是目前上皮性卵巢癌（EOC）标准治疗的一部分。然而，化疗耐药性常常迅速出现，导致化疗失败和预后不良。越来越多的证据表明，致癌长链非编码RNA H19在包括EOC在内的多种癌症中发挥重要作用。目前尚无研究探讨lncRNA-H19在EOC顺铂耐药中的作用及其潜在机制。通过qRT-PCR检测lncRNA-H19、miR-29b-3p和STAT3 mRNA的水平。用细胞计数试剂盒-8（CCK8）检测50%抑制浓度值。采用集落形成和CCK8试验检测细胞活力。用Transwells评估细胞迁移和侵袭能力。利用蛋白质免疫印迹法检测P-糖蛋白、多药耐药相关蛋白1（MRP1）、肺耐药相关蛋白（LRP）和STAT3蛋白水平。通过双荧光素酶、RNA免疫沉淀和RNA下拉实验验证lncRNA-H19与miR-29b-3p以及miR-29b-3p与STAT3之间的靶向关系。在顺铂耐药的EOC中，lncRNA-H19和STAT3显著升高，而miR-29b-3p降低。在化疗耐药的EOC细胞中，lncRNA-H19沉默可增强顺铂疗效。lncRNA-H19作为miR-29b-3p的竞争性内源性RNA，导致miR-29b-3p下游靶标STAT3的抑制解除，从而导致顺铂耐受的EOC产生化疗耐药。lncRNA-H19/miR-29b-3p轴通过靶向STAT3提高了EOC的顺铂耐药性，为改善EOC化疗提供了一种可能的方法。
