Zhou Yang, Wang Chunyan, Ding Jinye, Chen Yingying, Sun Yaoqi, Cheng Zhongping
Department of Gynecology and Obstetrics, Tenth People's Hospital, Tongji University School of Medicine, Shanghai, 200072, China.
Institute of Gynecological Minimally Invasive Surgery Research Center, Tongji University School of Medicine, Shanghai, 200072, China.
Cancer Cell Int. 2022 Jan 10;22(1):15. doi: 10.1186/s12935-021-02412-x.
Accumulating evidence has revealed that aberrant microRNA (miRNA) expression can affect the development of chemotherapy drug resistance by modulating the expression of relevant target proteins. Emerging evidence has demonstrated that miR-133a participates in the tumorigenesis of various cancers. However, whether miR-133a is associated with cisplatin resistance in ovarian cancer remains unclear.
To investigate the role of miR-133a in the development of cisplatin resistance in ovarian cancer.
MiR-133a expression in cisplatin-resistant ovarian cancer cell lines was assessed by reverse-transcription quantitative PCR (RT-qPCR). A cell counting kit-8 (CCK-8) assay was used to evaluate the viability of tumour cells treated with cisplatin in the presence or absence of miR-133a. A luciferase reporter assay was used to analyse the binding of miR-133a with the 3' untranslated region (3'UTR) of YES proto-oncogene 1 (YES1). The YES1 expression level was analysed using a dataset from the International Cancer Genome Consortium (ICGC) and assessed by RT-qPCR and western blotting in vitro. The roles and mechanisms of YES1 in cell functions were further probed via gain- and loss-of-function analysis.
The expression of miR-133a was significantly decreased in cisplatin-resistant ovarian cancer cell lines (A2780-DDP and SKOV3-DDP), and the overexpression of the miR-133a mimic reduced cisplatin resistance in A2780-DDP and SKOV3-DDP cells. Treatment with the miR-133a inhibitor increased cisplatin sensitivity in normal A2780 and SKOV3 cells. MiR-133a binds the 3'UTR of YES1 and downregulates its expression. Bioinformatics analysis revealed that YES1 expression was upregulated in recurrent cisplatin-resistant ovarian cancer tissue, and in vitro experiments also verified its upregulation in cisplatin-resistant cell lines. Furthermore, we discovered that miR-133a downregulated the expression of YES1 and thus inhibited cell autophagy to reduce cisplatin resistance. Yes1 knockdown significantly suppressed the cisplatin resistance of ovarian cancer cells by inhibiting autophagy in vitro. Xenograft tumour implantation further demonstrated that Yes1 overexpression promoted ovarian tumour development and cisplatin resistance.
Our results suggest that the miR-133a/YES1 axis plays a critical role in cisplatin resistance in human ovarian cancer by regulating cell autophagy, which might serve as a promising therapeutic target for ovarian cancer chemotherapy treatment in the future.
越来越多的证据表明，异常的微小RNA（miRNA）表达可通过调节相关靶蛋白的表达来影响化疗耐药性的发展。新出现的证据表明，miR-133a参与了多种癌症的肿瘤发生。然而，miR-133a是否与卵巢癌顺铂耐药相关仍不清楚。
探讨miR-133a在卵巢癌顺铂耐药发展中的作用。
采用逆转录定量PCR（RT-qPCR）检测顺铂耐药卵巢癌细胞系中miR-133a的表达。使用细胞计数试剂盒-8（CCK-8）试验评估在有或无miR-133a的情况下用顺铂处理的肿瘤细胞的活力。采用荧光素酶报告基因试验分析miR-133a与YES原癌基因1（YES1）的3'非翻译区（3'UTR）的结合。使用来自国际癌症基因组联盟（ICGC）的数据集分析YES1表达水平，并通过体外RT-qPCR和蛋白质免疫印迹法进行评估。通过功能获得和功能丧失分析进一步探讨YES1在细胞功能中的作用和机制。
miR-133a在顺铂耐药卵巢癌细胞系（A2780-DDP和SKOV3-DDP）中的表达显著降低，miR-133a模拟物的过表达降低了A2780-DDP和SKOV3-DDP细胞中的顺铂耐药性。用miR-133a抑制剂处理可增加正常A2780和SKOV3细胞对顺铂的敏感性。miR-133a与YES1的3'UTR结合并下调其表达。生物信息学分析显示，YES1表达在复发性顺铂耐药卵巢癌组织中上调，体外实验也证实其在顺铂耐药细胞系中上调。此外，我们发现miR-133a下调YES1的表达，从而抑制细胞自噬以降低顺铂耐药性。Yes1基因敲低通过在体外抑制自噬显著抑制了卵巢癌细胞的顺铂耐药性。异种移植瘤植入进一步证明Yes1过表达促进卵巢肿瘤发展和顺铂耐药。
我们的结果表明，miR-133a/YES1轴通过调节细胞自噬在人类卵巢癌顺铂耐药中起关键作用，这可能在未来作为卵巢癌化疗治疗的一个有前景的治疗靶点。
