MiR-382促进大鼠正常肝细胞BRL-3A增殖

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

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。

Objective To explore the effect of miR-382 on cell proliferation and apoptosis of rat hepatocyte line BRL-3 A in vitro. Methods BRL-3 A 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-3 A cells,and the expression of miR-382 was significantly down-regulated by treating with miR-382 inhibitor in the rat BRL-3 A cells（ P 0. 01）. MTT result showed that the activity of BRL-3 A 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-3 A 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. Conclusions MiR-382 may promote cell proliferation via regulating the expression of proliferation-related apoptosis-related proteins in rat hepatocyte line BRL-3 A.