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中国癌症防治杂志

中国癌症防治杂志 ›› 2022, Vol. 14 ›› Issue (1): 26-32.doi: 10.3969/j.issn.1674-5671.2022.01.05

• 基础研究 • 上一篇    下一篇

miR⁃145⁃5p靶向调控LOX基因对肝癌细胞侵袭和迁移的影响

  

  1. 广西医科大学附属肿瘤医院肝胆外科 
  • 出版日期:2022-02-25 发布日期:2022-03-11
  • 通讯作者: 齐鲁楠 E-mail：qilunan_gxmu@163.com
  • 基金资助:
    国家自然科学基金项目（81972306）；广西研究生教育创新计划学位与研究生教育改革课题（YCSW2020120）

 Effects of miR⁃145⁃5p on invasion and migration of liver cancer cells by targeting LOX gene

  • Online:2022-02-25 Published:2022-03-11

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摘要： 目的 探讨miR-145-5p靶向调控LOX对肝癌细胞侵袭和迁移的影响及其作用机制。方法 收集2017年9月至2019年9月广西医科大学附属肿瘤医院肝胆外科手术切除的73例肝癌患者肿瘤组织及其配对癌旁正常组织，以及肝癌细胞系SMMC-7721、SK-Hep1，采用RT-PCR法检测组织中miR-145-5p和LOX的表达。分别将miR-145-5p慢病毒载体、LOX过表达质粒及LOX慢病毒沉默载体转染至肝癌细胞SMMC-7721或SK-Hep1，同时用LOX过表达质粒和过表达miR-145-5p慢病毒载体共转染SMMC-7721细胞，各转染组均设置相应阴性对照组及空白对照组；然后采用Transwell实验检测肝癌细胞迁移和侵袭能力；Western blot法检测LOX蛋白的表达水平；通过TargetScan数据库预测miR-145-5p的靶点，并利用Cluster Profiler包进行GO和KEGG富集分析。结果 qRT-PCR检测结果显示，miR-145-5p在癌旁正常组织中的表达高于肝癌组织（4.196±2.288 vs 2.835±1.817，P<0.0001）；LOX在肝癌组织中的表达高于癌旁正常组织（12.17±1.369 vs 11.26±1.556，P<0.001）。Transwell检测结果显示，过表达LOX能促进肝癌细胞侵袭、迁移，敲低LOX则抑制肝癌细胞侵袭、迁移（均P<0.05）；过表达miR-145-5p能抑制肝癌细胞的侵袭、迁移能力，LOX过表达能逆转miR-145-5p对SMMC-7721细胞迁移和侵袭的抑制能力。TargetScan数据库预测显示，LOX是miR-145-5p的靶基因。Western blot检测结果显示，miR-145-5p能抑制LOX蛋白表达水平（P<0.001）。结论 miR-145-5p在肝癌组织中低表达，LOX则高表达，miR-145-5p可能通过负向调控LOX抑制肝癌细胞侵袭和迁移。

关键词: 肝癌, miR-145-5p, LOX, 侵袭, 迁移

Abstract: Objective  To investigate the effect of miR-145-5p targeting LOX on the invasion and migration of liver cancer cells and its mechanism. Methods The cancer tissues and adjacent normal tissues from 73 patients who underwent hepatobiliary surgery in Guangxi Medical University Cancer Hospital from September 2017 to September 2019, and liver cancer cell lines SMMC-7721 and SK-Hep1 were collected. The expressions of miR-145-5p and LOX in tissues were detected by RT-PCR. The miR-145-5p lentiviral vector, LOX overexpression plasmid and LOX lentiviral silencing vector were transfected with liver cancer cells SMMC-7721 or SK-Hep1; the LOX overexpressed plasmid and miR-145-5p lentivirus vector were co-transfected with SMMC-7721 cells; a corresponding negative control group and a blank control group were set in each transfection group. The migration and invasion ability of liver cancer cells were detected by Transwell assay; the expression level of LOX protein was detected by Western blot; The target of miR-145-5p was predicted by TargetScan database; and the GO and KEGG enrichment were analyzed by Cluster Profiler package. Results The results of qRT-PCR showed that miR-145-5p expression in adjacent normal tissues was higher than that in liver cancer tissues(4.196±2.288 vs 2.835±1.817, P<0.0001). The expression of LOX in liver cancer tissues was higher than that in adjacent normal tissues(12.17±1.369 vs 11.26±1.556, P<0.001). Transwell assay showed that the overexpression of LOX could promote the invasion and migration of liver cancer cells, while the knockdown of LOX inhibited the invasion and migration of liver cancer cells (all P<0.001). The overexpression of miR-145-5p could inhibit the invasion and migration of liver cancer cells, and the overexpression of LOX reversed the inhibitory ability of miR-145-5p on migration and invasion of SMMC-7721 cells. TargetScan database prediction showed that LOX was the target gene of miR-145-5p. Western blot showed that miR-145-5p could inhibit LOX protein expression (P<0.001). Conclusions miR-145-5p is lowly expressed in liver cancer tissues, while LOX is highly expressed. miR-145-5p may inhibit the invasion and migration of liver cancer cells by negatively regulating LOX.

Key words:  Liver cancer, miR-145-5p, LOX, Invasion, Migration

中图分类号: 

  • R735.7