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microRNA-383 通过下调 PARP2 抑制 PI3K-AKT-MTOR 信号通路抑制宫颈癌的发展。

microRNA-383 suppresses the PI3K-AKT-MTOR signaling pathway to inhibit development of cervical cancer via down-regulating PARP2.

机构信息

Department of Gynecology and Obstetrics, The Second Hospital of Shanxi Medical University, Taiyuan, P. R. China.

Department of Gynecology and Obstetrics, Xuzhou Central Hospital, Xuzhou, P. R. China.

出版信息

J Cell Biochem. 2018 Jul;119(7):5243-5252. doi: 10.1002/jcb.26585. Epub 2018 Mar 25.

Abstract

This study aims to evaluate the effect of the regulatory relationship between microRNA-383 (miR-383) and PARP2 in the cell migration and invasion in human with cervical cancer (CC) via the PI3K-AKT-MTOR signaling pathway. Cancerous tissues and corresponding paracancerous tissues were collected from 115 patients with CC. The positive expression rate of PARP2 was detected by immunohistochemistry. HeLa cells with highest miR-383 expression were selected and assigned into the blank, negative control (NC), miR-383 mimic, miR-383 inhibitor, si-PARP2, and miR-383 inhibitor + si-PARP2 groups. qRT-PCR and Western blot were performed to evaluate the expression of miR-383, PI3K, AKT, mTOR, PARP2, and p70S6K. MTT assay were utilized to measure cell viability. Transwell assay were applied to evaluate cell invasion and metastasis. Dual luciferase reporter assay identified that PARP2 is a target gene of miR-383. Cancerous tissues manifested higher expression of PI3K, AKT, mTOR, PARP2, and p70S6K but lower miR-383 expression than paracancerous tissues. Compared with the blank and NC groups, the miR-383 mimic and si-PARP2 groups had decreased expression of PI3K, AKT, mTOR, PARP2, and p70S6K mRNA and protein. In the miR-383 mimic and si-PARP2 groups, the cell viability, migration, and invasion were descended, in comparison to the blank and NC groups. All above parameters showed an opposite trend in the miR-383 inhibitor group when compared with the blank and NC groups. This study demonstrates that miR-383 could down-regulate PARP2 to protect against CC by inhibiting PI3K-AKT-MTOR signaling pathway.

摘要

本研究旨在通过 PI3K-AKT-MTOR 信号通路评估 microRNA-383(miR-383)与 PARP2 之间的调节关系对人宫颈癌(CC)细胞迁移和侵袭的影响。收集 115 例 CC 患者的癌组织及相应癌旁组织,采用免疫组织化学法检测 PARP2 阳性表达率。选取 miR-383 表达最高的 HeLa 细胞,分为空白组、阴性对照组(NC)、miR-383 模拟物组、miR-383 抑制剂组、si-PARP2 组和 miR-383 抑制剂+si-PARP2 组。qRT-PCR 和 Western blot 检测 miR-383、PI3K、AKT、mTOR、PARP2 和 p70S6K 的表达。MTT 法检测细胞活力。Transwell 法评估细胞侵袭和转移。双荧光素酶报告基因实验鉴定 PARP2 是 miR-383 的靶基因。癌组织中 PI3K、AKT、mTOR、PARP2 和 p70S6K 的表达均高于癌旁组织,miR-383 的表达均低于癌旁组织。与空白组和 NC 组相比,miR-383 模拟物组和 si-PARP2 组 PI3K、AKT、mTOR、PARP2 和 p70S6K mRNA 和蛋白表达均降低。与空白组和 NC 组相比,miR-383 模拟物组和 si-PARP2 组细胞活力、迁移和侵袭均降低,miR-383 抑制剂组与空白组和 NC 组比较,各参数均呈相反趋势。本研究表明,miR-383 可能通过抑制 PI3K-AKT-MTOR 信号通路下调 PARP2 来保护 CC。

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