Department of Thoracic Surgery, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, China.
Biosci Rep. 2020 Nov 27;40(11). doi: 10.1042/BSR20201858.
The present study aims to investigate the relationship between miR-19b-3p and esophageal cancer (ESCA), and to detect the effects of miR-19b-3p transferred by exosomes on the phenotype of EC9706 cells. The expression of miR-19b-3p was detected by starBase analysis and real-time quantitative PCR (RT-qPCR). The target genes of miR-19b-3p were predicted by TargetScan and further verified by luciferase analysis. The mRNA and protein expression levels of PTEN and EMT-related genes were detected by RT-qPCR and Western blotting. The effects of miR-19b-3p transferred by exosomes and its target genes on the apoptosis, migration and invasion of EC9706 cells were studied by establishing a co-culture model of donor cells. The expression of miR-19b-3p in ESCA plasma, cells and exosomes was significantly up-regulated. miR-19b-3p transferred by exosomes could significantly reduce EC9706 cells apoptosis rate, promote cell migration and invasion, and could target the inhibition of PTEN expression. PTEN overexpression promoted apoptosis, inhibited cell migration and invasion, down-regulated the expression of MMP-2 and vimentin, and up-regulated E-cadherin expression; however, these effects could be partially reversed by miR-19b-3p. In summary, our results reveal that miR-19b-3p transferred by exosomes can target PTEN to regulate ESCA biological functions in the receptor EC9706 cells.
本研究旨在探讨 miR-19b-3p 与食管癌（ESCA）之间的关系，并检测外泌体介导的 miR-19b-3p 对 EC9706 细胞表型的影响。通过 starBase 分析和实时定量 PCR（RT-qPCR）检测 miR-19b-3p 的表达。通过 TargetScan 预测 miR-19b-3p 的靶基因，并通过荧光素酶分析进一步验证。通过 RT-qPCR 和 Western blot 检测 PTEN 和 EMT 相关基因的 mRNA 和蛋白表达水平。通过建立供体细胞共培养模型，研究外泌体转染的 miR-19b-3p 及其靶基因对 EC9706 细胞凋亡、迁移和侵袭的影响。ESCA 血浆、细胞和外泌体中 miR-19b-3p 的表达明显上调。外泌体转染的 miR-19b-3p 可显著降低 EC9706 细胞凋亡率，促进细胞迁移和侵袭，并能靶向抑制 PTEN 表达。PTEN 过表达促进细胞凋亡，抑制细胞迁移和侵袭，下调 MMP-2 和波形蛋白表达，上调 E-钙黏蛋白表达；然而，这些作用可被 miR-19b-3p 部分逆转。综上所述，本研究结果表明，外泌体转染的 miR-19b-3p 可以靶向 PTEN 调节受体 EC9706 细胞中的 ESCA 生物学功能。
