Gao Xiaojie, Jiang Yan, Li Yingying
Department of Pharmacy, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou Zhejiang 310005, P.R. China.
Department of Pharmacy, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310006, P.R. China.
Exp Ther Med. 2021 May;21(5):507. doi: 10.3892/etm.2021.9938. Epub 2021 Mar 18.
To investigate the role of microRNA (miR)-140-5p in doxorubicin (DOX) sensitivity in hepatocellular carcinoma, miR-140-5p and peptidyl-prolyl isomerase NIMA-interacting 1 (PIN1) expression was first evaluated in hepatocellular carcinoma tissues using starBase. Next, experiments were performed. Cell line expression of miR-140-5p and PIN1 expression was detected by reverse transcription polymerase chain reaction. Cell viability and proliferation were determined by the Cell Counting Kit-8 and EdU assays. The relationship between miR-140-5p and PIN1 was evaluated by TargetScan and a luciferase reporter system. Western blotting was used to detect the expression of PIN1. It was observed that miR-140-5p was downregulated in hepatocellular carcinoma tissues and cell lines compared with normal samples in HCC or normal liver cells. Gain-of-function experiments revealed that miR-140-5p mimics were able to enhance DOX sensitivity of hepatocellular carcinoma cells. Further studies revealed that PIN1 was a target gene of miR-140-5p. Suppression of PIN1 led to higher DOX sensitivity in hepatocellular carcinoma cells. Finally, when comparing a PIN1-siRNA alone group and a PIN1-siRNA plus miR-140-5p inhibitor group, there was no significant difference in cell viability. Furthermore, miR-140-5p mimics did not reduce the sensitivity of PIN1mut plasmid to DOX in HUH7 and SNU449 cells. The present study demonstrated that miR-140-5p could enhance DOX sensitivity in hepatocellular carcinoma cells by targeting PIN1.
为研究微小RNA（miR）-140-5p在肝细胞癌对多柔比星（DOX）敏感性中的作用，首先使用starBase评估肝细胞癌组织中miR-140-5p和肽基脯氨酰异构酶NIMA相互作用蛋白1（PIN1）的表达。接下来，进行实验。通过逆转录聚合酶链反应检测细胞系中miR-140-5p和PIN1的表达。采用细胞计数试剂盒-8和EdU检测法测定细胞活力和增殖情况。通过TargetScan和荧光素酶报告系统评估miR-140-5p与PIN1之间的关系。采用蛋白质免疫印迹法检测PIN1的表达。结果观察到，与肝癌或正常肝细胞中的正常样本相比，miR-140-5p在肝细胞癌组织和细胞系中表达下调。功能获得实验表明，miR-140-5p模拟物能够增强肝癌细胞对DOX的敏感性。进一步研究表明，PIN1是miR-140-5p的靶基因。抑制PIN1可使肝癌细胞对DOX的敏感性更高。最后，比较单独使用PIN1-siRNA组和PIN1-siRNA加miR-140-5p抑制剂组时，细胞活力无显著差异。此外，miR-140-5p模拟物并未降低HUH7和SNU449细胞中PIN1mut质粒对DOX的敏感性。本研究表明，miR-140-5p可通过靶向PIN1增强肝癌细胞对DOX的敏感性。
