微RNA-490-5p抑制人肝内胆管细胞癌RBE细胞的侵袭和迁移

发布时间：2018-05-31 11:17

本文选题：肝内胆管癌 + 微RNAs　；参考：《肿瘤》2017年07期

【摘要】：目的:研究微RNA-490-5p(microRNA-490-5p,miR-490-5p)在人肝内胆管细胞癌组织中的表达情况,并探讨其对肝内胆管癌RBE细胞侵袭和迁移能力的影响及可能的分子机制。方法:采用实时荧光定量PCR法检测miR-490-5p在5对肝内胆管细胞癌组织及其癌旁正常组织中的表达水平。利用脂质体瞬时转染法将miR-490-5p模拟物及其阴性对照转入人肝内胆管细胞癌RBE细胞中。随后,采用划痕愈合实验以及Transwell小室迁移实验检测RBE细胞迁移能力的变化,Transwell小室侵袭实验检测RBE细胞侵袭能力的变化;最后,通过CCK-8法、平板克隆形成实验和FCM法检测RBE细胞增殖及凋亡的变化。应用在线miRNA靶基因预测软件预测miR-490-5p的靶基因,并采用蛋白印迹法对预测的靶基因进行验证。结果:miR-490-5p在人肝内胆管细胞癌组织中明显下调(P0.05)。与阴性对照组相比,miR-490-5p模拟物转染组RBE细胞的增殖、克隆形成能力和凋亡均无明显变化(P值均0.05);划痕愈合实验以及Transwell小室迁移实验检测结果显示,RBE细胞的迁移能力被明显抑制(P值均0.05);Transwell小室侵袭实验检测结果显示,RBE细胞的侵袭能力被明显抑制(P值0.05)。Target Scan网站预测转录因子FBJ骨肉瘤癌基因(FBJ osteosarcoma oncogene,c-Fos)可能是miR-490-5p的潜在靶基因;蛋白质印迹法证实,miR-490-5p模拟物转染组细胞中c-Fos蛋白的表达水平明显下调(P0.05)。结论:miR-490-5p在人肝内胆管细胞癌组织中低表达,而且其可能通过下调c-Fos的表达来调控人肝内胆管细胞癌RBE细胞的迁移和侵袭。
[Abstract]:Aim: to investigate the expression of microRNA-490-5pnmmiR-490-5p in human intrahepatic cholangiocarcinoma and its effect on the invasion and migration of RBE cells and its possible molecular mechanism. Methods: the expression of miR-490-5p in 5 pairs of intrahepatic cholangiocarcinoma and its adjacent normal tissues was detected by real-time fluorescence quantitative PCR. MiR-490-5p mimics and their negative controls were transfected into human intrahepatic cholangiocarcinoma RBE cells by liposome transient transfection. Then, the change of migration ability of RBE cells was detected by scratch healing test and Transwell chamber migration test. Finally, the change of RBE cell invasion ability was detected by CCK-8 assay. The proliferation and apoptosis of RBE cells were detected by plate clone formation assay and FCM assay. The target gene of miR-490-5p was predicted by online miRNA target gene prediction software, and the predicted target gene was verified by Western blotting. Results: 1 miR-490-5p significantly down-regulated P0.05 in human intrahepatic cholangiocarcinoma. Compared with the negative control group, the proliferation of RBE cells was observed in the RBE cells transfected with mimics of miR-490-5p. The results of scratch healing test and Transwell chamber migration test showed that the migration ability of Transwell cells was significantly inhibited. The ability of cell invasion was significantly inhibited by 0.05).Target Scan website. It was predicted that the transcription factor FBJ osteosarcoma oncogene (FBJ osteosarcoma oncogene c-Fos) might be a potential target gene of miR-490-5p. Western blotting showed that the expression level of c-Fos protein was significantly down-regulated in the cells transfected with the mimic of miR-490-5p. Conclusion the low expression of 1% miR-490-5p in human intrahepatic cholangiocarcinoma may regulate the migration and invasion of human intrahepatic cholangiocarcinoma RBE cells by down-regulating the expression of c-Fos.
【作者单位】：上海交通大学医学院附属新华医院普外科;上海市胆道疾病研究中心;上海交通大学医学院胆道疾病研究所;
【基金】：国家自然科学基金资助项目(编号:81272402)~~
【分类号】：R735.8

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