MiR-382促进大鼠正常肝细胞BRL-3A增殖

高航 张春艳 王凤娟 徐存拴*

doi:10.16098/j.issn.0529-1356.2018.04.004

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

细胞和分子生物学

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*

作者信息 +

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

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

Author information +

文章历史 +

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。

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.

导出引用

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 https://doi.org/10.16098/j.issn.0529-1356.2018.04.004

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 https://doi.org/10.16098/j.issn.0529-1356.2018.04.004

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