Department of Gynaecology, Shanghai University of Medicine & Health Sciences Affiliated Zhoupu Hospital, Shanghai, China.
Department of Gynaecology, Obstetrics and Gynaecology Hospital, Fudan University, Shanghai, China; Department of Obstetrics and Gynaecology of Shanghai Medical School, Fudan University, Shanghai, China; Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Shanghai, China.
Pathol Res Pract. 2020 Jul;216(7):152979. doi: 10.1016/j.prp.2020.152979. Epub 2020 Apr 28.
miR-194-5p has been associated with drug resistance in many cancers. However, the role of miR-194-5p in cisplatin resistance in ovarian cancer is still unclear.
To study the role and mechanism of miR-194-5p in cisplatin resistance, qRT-PCR was performed to determine the expression of miR-194-5p and SLC40A1 in ovarian cancer. Cell Counting Kit-8 (CCK8) assay was used to analyse cell viability after cisplatin treatment. Dual-luciferase reporter gene assay was performed to examine the relationship between miR-194-5p and SLC40A1. The genes downstream of SLC40A1 were investigated through bioinformatics analysis.
Compared to cisplatin-sensitive ovarian cancer cells, higher miR-194-5p expression and lower SLC40A1 expression were found in cisplatin-resistant ovarian cancer cells. Moreover, this study demonstrated that over-expression of miR-194-5p inhibited SLC40A1 expression, and knockdown of miR-194-5p promoted SLC40A1 expression. In addition, dual-luciferase reporter gene assay further confirmed the negative correlation between miR-194-5p and SLC40A1. Furthermore, we found that over-expression of miR-194-5p resulted in cisplatin resistance. When miR-194-5p and SLC40A1 were simultaneously up-regulated, cisplatin sensitivity increased, while down-regulation of miR-194-5p sensitised ovarian cancer cells to cisplatin. However, when miR-194-5p and SLC40A1 were simultaneously down-regulated, cisplatin sensitivity was decreased. These data suggested that miR-194-5p inhibited SLC40A1 expression to induce cisplatin resistance. In addition, bioinformatics analysis indicated a positive correlation of SLC40A1 with hephaestin (HEPH), and homeostatic iron regulator (HFE). However, we found that HEPH and HFE were associated with cisplatin resistance, suggesting that their role in drug resistance is induced by miR-194-5p/SLC40A1.
In conclusion, we found that miR-194-5p inhibited SLC40A1 expression to induce cisplatin resistance in ovarian cancer. This study suggests that miR-194-5p could be a potential therapeutic target and a prognostic biomarker for ovarian cancer, with important implications for future research.
miR-194-5p 与许多癌症的耐药性有关。然而，miR-194-5p 在卵巢癌顺铂耐药中的作用尚不清楚。
为了研究 miR-194-5p 在顺铂耐药中的作用和机制，通过 qRT-PCR 测定卵巢癌细胞中 miR-194-5p 和 SLC40A1 的表达。细胞计数试剂盒-8（CCK8）检测顺铂处理后细胞活力。双荧光素酶报告基因检测 miR-194-5p 与 SLC40A1 之间的关系。通过生物信息学分析研究 SLC40A1 下游的基因。
与顺铂敏感的卵巢癌细胞相比，顺铂耐药的卵巢癌细胞中 miR-194-5p 表达升高，SLC40A1 表达降低。此外，本研究表明过表达 miR-194-5p 抑制 SLC40A1 表达，而敲低 miR-194-5p 促进 SLC40A1 表达。此外，双荧光素酶报告基因检测进一步证实了 miR-194-5p 与 SLC40A1 之间的负相关关系。此外，我们发现过表达 miR-194-5p 导致顺铂耐药。当 miR-194-5p 和 SLC40A1 同时上调时，顺铂敏感性增加，而下调 miR-194-5p 可使卵巢癌细胞对顺铂敏感。然而，当 miR-194-5p 和 SLC40A1 同时下调时，顺铂敏感性降低。这些数据表明 miR-194-5p 通过抑制 SLC40A1 表达诱导顺铂耐药。此外，生物信息学分析表明 SLC40A1 与 hephaestin（HEPH）和铁稳态调节因子（HFE）呈正相关。然而，我们发现 HEPH 和 HFE 与顺铂耐药有关，这表明它们在耐药性中的作用是由 miR-194-5p/SLC40A1 诱导的。
综上所述，我们发现 miR-194-5p 通过抑制 SLC40A1 表达诱导卵巢癌顺铂耐药。本研究表明 miR-194-5p 可能是卵巢癌潜在的治疗靶点和预后生物标志物，对未来的研究具有重要意义。
