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[微小RNA let-7g-3p通过靶向HMGB2调控膀胱癌细胞的增殖、迁移、侵袭和凋亡]

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

作者信息

Zou Z, Cheng Q, Li Z, Gao W, Sun W, Liu B, Guo Y, Liu J

机构信息

Department of Urology, First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2022 Sep 20;42(9):1335-1343. doi: 10.12122/j.issn.1673-4254.2022.09.09.

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 bioinformatics analysis, 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 ( < 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 ( < 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.

摘要

目的

探讨微小RNA let-7g-3p调控膀胱癌细胞生物学行为的分子机制。

方法

采用qRT-PCR检测let-7g-3p在膀胱癌组织及癌旁组织、正常膀胱上皮细胞(HUC细胞)和膀胱癌细胞(T24、5637和EJ细胞)中的表达水平。将let-7g-3p模拟物或抑制剂转染至T24细胞,检测细胞增殖、迁移、侵袭及凋亡的变化。对过表达let-7g-3p的细胞进行转录组测序,生物信息学分析、双荧光素酶报告基因检测、qRT-PCR及蛋白质免疫印迹结果证实HMGB2基因是let-7g-3p的靶基因。检测HMGB2在HUC、T24、5637和EJ细胞中的表达情况,在敲低HMGB2基因的细胞中观察敲低let-7g-3p对生物学行为的影响。

结果

qRT-PCR证实let-7g-3p在膀胱癌组织和细胞中的表达显著降低(<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,而敲低let-7g-3p则产生相反的作用。生物信息学和转录组测序结果表明,HMGB2是介导let-7g-3p对膀胱癌细胞作用的关键分子。荧光素酶报告基因检测、qRT-PCR及蛋白质免疫印迹均证实let-7g-3p对HMGB2具有负调控作用(<0.01)。敲低HMGB2可部分逆转敲低let-7g-3p对膀胱癌细胞生物学行为的影响。

结论

微小RNA let-7g-3p可通过负调控HMGB2基因抑制膀胱癌细胞的生物学行为。

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