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microRNA-142-5p 通过靶向磷酸酶张力蛋白同源物调节乳腺癌细胞增殖和凋亡。

MicroRNA‑142‑5p modulates breast cancer cell proliferation and apoptosis by targeting phosphatase and tensin homolog.

机构信息

Department of General Surgery, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, P.R. China.

出版信息

Mol Med Rep. 2018 Jun;17(6):7529-7536. doi: 10.3892/mmr.2018.8812. Epub 2018 Mar 28.

Abstract

A total of 60 breast cancer (BC) tissues and adjacent healthy tissues from patients who underwent surgery in Renmin Hospital of Wuhan University were collected for analysis in the present study. Results from reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) demonstrated that, compared with the adjacent healthy tissues, the expression levels of microRNA (miR)‑142‑5p were significantly elevated in BC tissues. Bioinformatics analysis was performed using TargetScan for the prediction of potential target sites that matched the seed region of miR‑142‑5p; phosphatase and tensin homolog (PTEN) exhibited the highest score and was selected for further analysis. Results of RT‑qPCR analysis demonstrated that, compared with the adjacent healthy tissues, the mRNA expression levels of PTEN were significantly decreased in breast cancer tissues. miR‑142‑5p and PTEN expression levels were positively and negatively associated, respectively, with patient tumor size and metastasis. MDA‑MB‑231 cells were divided into three groups including the Control group, the miR‑NC inhibitor group and the miR‑142‑5p inhibitor group. As for alterations in cell behavior, including cell viability and cell apoptosis, and protein expression levels, there were no significant differences between Control and miR‑NC inhibitor groups. MTT assay results revealed that, compared with Control and miR‑NC inhibitor groups, miR‑142‑5p inhibitor reduced MDA‑MB‑231 cell proliferation. Flow cytometric analysis demonstrated that, compared with Control and miR‑NC inhibitor groups, miR‑142‑5p inhibitor treatment induced MDA‑MB‑231 cell apoptosis. Western blotting results demonstrated that, compared with Control and miR‑NC inhibitor groups, miR‑142‑5p inhibitor treatment significantly increased the expression of PTEN, reduced the activation of phosphatidylinositol‑4,5‑bisphosphate 3‑kinase/RACα serine/threonine‑protein kinase signaling. Finally, PTEN was demonstrated to interact with miR‑142‑5p from the results of dual‑luciferase reporter assay in the present study. The findings of the present study suggested that miR‑142‑5p may be a potential therapeutic target for the future investigations and insights for breast cancer.

摘要

本研究收集了 60 例武汉人民医院手术患者的乳腺癌(BC)组织和相邻的健康组织进行分析。逆转录-定量聚合酶链反应(RT-qPCR)的结果表明,与相邻的健康组织相比,BC 组织中 microRNA(miR)-142-5p 的表达水平显著升高。使用 TargetScan 进行生物信息学分析,以预测与 miR-142-5p 种子区匹配的潜在靶位点;磷酸酶和张力蛋白同源物(PTEN)得分最高,并被选择用于进一步分析。RT-qPCR 分析结果表明,与相邻的健康组织相比,乳腺癌组织中 PTEN 的 mRNA 表达水平显著降低。miR-142-5p 和 PTEN 的表达水平分别与患者肿瘤大小和转移呈正相关和负相关。MDA-MB-231 细胞分为三组:对照组、miR-NC 抑制剂组和 miR-142-5p 抑制剂组。关于细胞行为的改变,包括细胞活力和细胞凋亡以及蛋白表达水平,对照组和 miR-NC 抑制剂组之间没有显著差异。MTT 检测结果表明,与对照组和 miR-NC 抑制剂组相比,miR-142-5p 抑制剂降低了 MDA-MB-231 细胞的增殖。流式细胞术分析表明,与对照组和 miR-NC 抑制剂组相比,miR-142-5p 抑制剂处理诱导 MDA-MB-231 细胞凋亡。Western blot 结果表明,与对照组和 miR-NC 抑制剂组相比,miR-142-5p 抑制剂处理显著增加了 PTEN 的表达,降低了磷脂酰肌醇-4,5-二磷酸 3-激酶/RACα 丝氨酸/苏氨酸-蛋白激酶信号的激活。最后,通过本研究的双荧光素酶报告基因检测证实了 PTEN 与 miR-142-5p 相互作用。本研究的结果表明,miR-142-5p 可能是未来乳腺癌研究和见解的一个潜在治疗靶点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bde3/5983952/3cbe3e4490c4/MMR-17-06-7529-g00.jpg

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