南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (9): 1335-1343.doi: 10.12122/j.issn.1673-4254.2022.09.09

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MiR-let-7g-3p靶向HMGB2调控膀胱癌细胞的增殖、迁移、侵袭和凋亡

邹震海，程 琪，李 中，高五岳，孙 巍，刘贝贝，郭园园，刘建民   

  1. 蚌埠医学院第一附属医院泌尿外科，安徽 蚌埠 233004
  • 出版日期:2022-09-20 发布日期:2022-09-28

microRNA let-7g-3p regulates proliferation, migration, invasion and apoptosis of bladder cancer cells by targeting HMGB2

ZOU Zhenhai, CHENG Qi, LI Zhong, GAO Wuyue, SUN Wei, LIU Beibei, GUO Yuanyuan，LIU Jianmin   

  1. Department of Urology, First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, China
  • Online:2022-09-20 Published:2022-09-28

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摘要： 目的 探讨MiR-let-7g-3p靶向HMGB2对膀胱癌细胞生物学行为的影响及其分子机制。方法 通过qRT-PCR检测膀胱癌组织和癌旁组织以及正常尿路上皮细胞SV-HUC-1和膀胱癌细胞（T24、5637和EJ）中let-7g-3p的表达水平；通过转染过表达或敲低T24细胞let-7g-3p表达，检测细胞增殖、迁移和侵袭、和凋亡等变化；对过表达let-7g-3p的细胞进行转录组测序，结合生物信息学分析，通过双荧光素酶报告实验、qRT-PCR和Western-blot分析let-7g-3p靶向负性调控HMGB2基因；qRT-PCR检测正常尿路上细胞和膀胱癌细胞（T24、5637和EJ）中HMGB2的表达水平，并且在敲低HMGB2的细胞株中，敲低let-7g-3p，检测细胞生物学行为改变。结果 qRT-qPCR证实let-7g-3p在膀胱癌组织和细胞表达明显下调（P<0.01）；过表达let-7g-3p可抑制细胞增殖、迁移和侵袭，促进细胞凋亡（P<0.01）；相反，敲减let-7g-3p表达可促进细胞增殖、迁移和侵袭，抑制细胞凋亡（P<0.01）。生物信息学和转录组测序结果显示，HMGB2是let-7g-3p作用于膀胱癌细胞的关键分子；双荧光素酶报告实验、qRT-PCR和Westernblot显示，HMGB2受let-7g-3p的负性调控；HMGB2膀胱癌细胞中表达水平均明显上调（P<0.01），敲低HMGB2可部分逆转敲低let-7g-3p对膀胱癌细胞生长的促进作用。结论 MiRNA-let-7g-3p通过靶向负性调控HMGB2基因抑制膀胱癌细胞的生物行为。

关键词: let-7g-3p；HMGB2基因；膀胱肿瘤

Abstract: Objective To explore the molecular mechanism by which microRNA let-7g-3p regulates biological behaviors of bladder cancer cells. Methods The expression levels of let-7g-3p in bladder cancer and adjacent tissues, normal bladder epithelial cells (HUC cells) and bladder cancer cells (T24, 5637 and EJ cells) were detected using qRT- PCR. T24 cells were transfected with let-7g-3p mimic or inhibitor, and the changes in cell proliferation, migration, invasion, and apoptosis were examined. Transcriptome sequencing was carried out in cells overexpressing let-7g-3p, and the results of bioinformaticsanalysis, double luciferase reporter gene assay, qRT- PCR and Western blotting confirmed that HMGB2 gene was the target gene of let-7g-3p. The expression of HMGB2 was examined in HUC, T24, 5637 and EJ cells, and in cells with HMGB2 knockdown, the effect of let-7g-3p knockdown on the biological behaviors were observed. Results qRT-qPCR confirmed that let-7g-3p expression was significantly lower in bladder cancer tissues and cells (P<0.01). Overexpression of let-7g-3p inhibited cell proliferation, migration and invasion, and promoted cell apoptosis, while let-7g-3p knock-down produced the opposite effects. Bioinformatics and transcriptome sequencing results showed that HMGB2 was the key molecule that mediate the effect of let-7g-3p on bladder cancer cells. Luciferase reporter gene assay, qRT-PCR and Western blotting all confirmed that HMGB2 was negatively regulated by let-7g-3p (P<0.01). Knocking down HMGB2 could partially reverse the effect of let-7g-3p knockdown on the biological behaviors of the bladder cancer cells. Conclusion The microRNA let-7g-3p can inhibit the biological behavior of bladder cancer cells by negatively regulating HMGB2 gene.

Key words: let-7g-3p; HMGB2; bladder cancer