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微小RNA-21通过靶向磷脂酰肌醇-3-激酶/蛋白激酶B通路促进肝星状细胞增殖并抑制其凋亡。

miR-21 promotes proliferation and inhibits apoptosis of hepatic stellate cells through targeting PTEN/PI3K/AKT pathway.

作者信息

Hao Xin-Jie, Xu Cheng-Zhen, Wang Jin-Tai, Li Xiao-Jie, Wang Ming-Min, Gu Yi-Hai, Liang Zhi-Gang

机构信息

a Hepatic Department , No. 6 Qingdao People's Hospital , Qingdao , China.

b Orthopedics Department , No. 8 Qingdao People's Hospital , Qingdao , China.

出版信息

J Recept Signal Transduct Res. 2018 Oct-Dec;38(5-6):455-461. doi: 10.1080/10799893.2019.1585452.

Abstract

To investigate the effect of microRNA 21 (miR-21) on hepatic stellate cells (HSCs) proliferation and apoptosis, and further to study its potential mechanisms. LX-2 cells were divided into miR-21 mimic group (Mimic), miR-21 mimic negative control group (NM), miR-21 inhibitor group (Inhibitor), miR-21 inhibitor negative control group (NC), and blank control group (Control). The cell proliferation was detected by CCK-8 assay and the cell migration and invasion were detected by scratch and transwell assay. Cell cycle and apoptosis were detected by flow cytometry. The levels of interleukin (IL)-6, tumor necrosis factor (TNF)-α, and transforming growth factor (TGF)-β1 were detected by enzyme-linked immunosorbent assay (ELISA). Proliferation, apoptosis, and phosphatase and tensin homolog (PTEN)/phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway related genes and proteins were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot, respectively. The cells proliferation, migration, and invasion were promoted in Mimic group. The levels of IL-6, TNF-α, and TGF-β1 were increased after miR-21 administration. The expression of α-smooth muscle actin (SMA) and collagen 1 (Colla1) were increased, while Bax/B-cell lymphoma (Bcl)-2 ratio and programed cell death 4 (PDCD4) were reduced after miR‑21 treatment. Meanwhile, the mRNA and protein expression of PTEN were reduced and PI3K/AKT pathway been promoted. Our study demonstrated that miR-21 could promote proliferation and inhibit apoptosis of HSCs, and its mechanism may be related to PTEN/PI3K/AKT pathway.

摘要

为研究微小RNA 21(miR-21)对肝星状细胞(HSCs)增殖和凋亡的影响,并进一步探讨其潜在机制。将LX-2细胞分为miR-21模拟物组(模拟物组)、miR-21模拟物阴性对照组(NM组)、miR-21抑制剂组(抑制剂组)、miR-21抑制剂阴性对照组(NC组)和空白对照组(对照组)。采用CCK-8法检测细胞增殖,划痕法和Transwell法检测细胞迁移和侵袭。通过流式细胞术检测细胞周期和凋亡。采用酶联免疫吸附测定(ELISA)检测白细胞介素(IL)-6、肿瘤坏死因子(TNF)-α和转化生长因子(TGF)-β1水平。分别采用定量实时聚合酶链反应(qRT-PCR)和蛋白质印迹法检测增殖、凋亡以及磷酸酶和张力蛋白同源物(PTEN)/磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)信号通路相关基因和蛋白。模拟物组细胞的增殖、迁移和侵袭均增强。给予miR-21后,IL-6、TNF-α和TGF-β1水平升高。miR-21处理后,α平滑肌肌动蛋白(SMA)和胶原蛋白1(Colla1)表达增加,而Bax/B细胞淋巴瘤(Bcl)-2比值和程序性细胞死亡4(PDCD4)降低。同时,PTEN的mRNA和蛋白表达降低,PI3K/AKT通路被激活。我们的研究表明,miR-21可促进肝星状细胞增殖并抑制其凋亡,其机制可能与PTEN/PI3K/AKT通路有关。

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