MiR-382促进大鼠正常肝细胞BRL-3A增殖

doi:10.16098/j.issn.0529-1356.2018.04.004

解剖学报 ›› 2018, Vol. 49 ›› Issue (4): 437-442.doi: 10.16098/j.issn.0529-1356.2018.04.004

MiR-382促进大鼠正常肝细胞BRL-3A增殖

高航¹ 张春艳¹ 王凤娟² 徐存拴^1*

1. 河南师范大学生命科学学院，河南省-科技部共建细胞分化调控国家重点实验室培育基地，河南新乡453007；2. 太康县第一高级中学，河南太康461400

收稿日期:2017-10-25 修回日期:2018-01-06 出版日期:2018-08-06 发布日期:2018-08-06
通讯作者: 徐存拴 E-mail:cellkeylab@126.com
基金资助:
4种共表达microRNA协同调节大鼠肝细胞增殖和肝再生的分子机制研究

MiR-382 promoting BRL-3A cell proliferation of rat

GAO Hang¹ ZHANG Chun-yan¹ WHANG Feng-juan² XU Cun-shuan^1*

1. State Key Laboratory Cultivation Base for Cell Differentiation Regulation, College of Life Science, Henan Normal University, He’nan Xinxiang453007, China; 2.Taikang First Senior High School, He’nan Taikang461400, China

Received:2017-10-25 Revised:2018-01-06 Online:2018-08-06 Published:2018-08-06
Contact: XU Cun-shuan E-mail:cellkeylab@126.com

摘要/Abstract

摘要：

目的探讨miR-382对大鼠正常肝细胞BRL-3A增殖和凋亡的影响。方法 BRL-3A细胞瞬时转染miR-382的模拟物和抑制物48 h， MTT法测定细胞活性；流式细胞术检测细胞周期；Real-time PCR检测细胞增殖和凋亡相关基因的表达情况。结果在大鼠BRL-3A细胞中，转染模拟物可以显著增加miR-382的表达，转染抑制物可以显著降低miR-382的表达(P＜0.01)。MTT检测表明，miR-382过表达后，BRL-3A细胞活性明显提高；流式细胞术检测表明，miR-382过表达组S期的细胞数明显增加，而miR-382干涉组S期的细胞数明显减少；用Real-time PCR检测细胞增殖/凋亡相关基因mRNA表达水平表明，miR-382过表达后，BRL-3A细胞增殖相关基因增殖细胞核抗原（PCNA)、Bcl-2和Ccnd1的mRNA表达水平上调，细胞凋亡相关基因Caspase-3和Bax的mRNA表达水平下调，而miR-382干涉组与上述结果相反。结论 miR-382通过促进细胞增殖相关基因表达，抑制细胞凋亡相关基因表达而促进大鼠正常肝细胞BRL-3A增殖。

关键词: MiR-382, BRL-3A细胞, 细胞周期, 细胞增殖, 实时定量聚合酶链反应, 大鼠

Abstract:

Objective To explore the effect of miR-382 on cell proliferation and apoptosis of rat hepatocyte line BRL-3A in vitro. Methods BRL-3A cells were transiently transfected with miR-382 mimics and inhibitors for 48 hours，MTT assay was used to detect the cell viability. Flow cytometry was used to observe the cell cycle. The expression of proliferation/apoptosis-related genes was detected by Real-time PCR. Results The expression of miR-382 was significantly up-regulated by treating with miR-382 mimics in the rat BRL-3A cells，and the expression of miR-382 was significantly down-regulated by treating with miR-382 inhibitor in the rat BRL-3A cells(P<0.01). MTT result showed that the activity of BRL-3A cells was significantly increased after miR-382 overexpression. Flow cytometry showed that the number of cells in S phase of miR-382 overexpression group was significantly higher than that of the negative control group, while the number of cells in S phase of miR-382 interference group was significantly lower than that of the negative control group. The mRNA expression levels of poliferating cell nuclear antigen(PCNA), Bcl-2 and Ccnd1 in BRL-3A cells was up-regulated and the mRNA expression levels of Caspase-3 and Bax were down-regulated by Real-time PCR after miR-382 overexpression, while the miR-382 interference group was opposite to the above result. Conclusion sMiR-382 may promote cell proliferation via regulating the expression of proliferation-related apoptosis-related proteins in rat hepatocyte line BRL-3A.

Key words: MiR-382, BRL-3A cell, Cell cycle, Cell proliferation, Real-time PCR, Rat

高航张春艳王凤娟徐存拴. MiR-382促进大鼠正常肝细胞BRL-3A增殖[J]. 解剖学报, 2018, 49(4): 437-442.

GAO Hang ZHANG Chun-yan WHANG Feng-juan XU Cun-shuan. MiR-382 promoting BRL-3A cell proliferation of rat[J]. Acta Anatomica Sinica, 2018, 49(4): 437-442.

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MiR-382促进大鼠正常肝细胞BRL-3A增殖

MiR-382 promoting BRL-3A cell proliferation of rat

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