新蛋白C7orf42促进大鼠肝细胞BRL-3A增殖

新蛋白C7orf42促进大鼠肝细胞BRL-3A增殖

张春艳常翠芳张世馥马纪张富春徐存拴

解剖学报 ›› 2017, Vol. 48 ›› Issue (2) : 150-155.

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解剖学报 ›› 2017, Vol. 48 ›› Issue (2) : 150-155. DOI: 10.16098/j.issn.0529-1356.2017.02.005

细胞和分子生物学

新蛋白C7orf42促进大鼠肝细胞BRL-3A增殖

张春艳^1,2常翠芳² 张世馥²马纪¹张富春¹ 徐存拴^2*

作者信息 +

Novel protein C7orf42 promotes rat hepatocyte BRL-3A proliferation

ZHANG Chun-yan^1,2 CHANG Cui-fang²ZHANG Shi-fu²MA Ji¹ZHANG Fu-chun¹XU Cun-shuan^2*

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文章历史 +

摘要

目的探讨未知功能蛋白C7orf42对体外培养的大鼠肝细胞BRL-3A增殖的影响。方法基因干涉下调C7orf42表达后，用MTT、EdU掺入法检测C7orf42对BRL-3A细胞增殖的影响，流式细胞术检测细胞周期进程以及Reat-time PCR和Western blotting检测细胞增殖相关基因表达。结果 MTT法检测表明，转染C7orf42干涉片段后48h，干涉组比阴性对照组细胞活力降低11%（P<0.05）；EdU掺入法检测表明，实验组的EdU阳性细胞比率比对照组降低了13%（P<0.05）；流式细胞术检测表明，干涉组比阴性对照组S+G2/M期的细胞数降低12%（P<0.05）；Reat-time PCR检测表明，干涉C7orf42表达后细胞增殖相关基因JUN、CCND1、MYC和CCNA2的表达分别下调26%、31%、37%和14%（P<0.05）；Western blotting检测表明，干涉C7orf42表达后细胞增殖相关蛋白JUN、CCND1、MYC和CCNA2的表达分别下调59%、54%、18%和27%（P<0.05）。结论 C7orf42可能通过上调细胞增殖相关蛋白JUN、CCND1、MYC和CCNA2的表达，促进体外培养的大鼠肝细胞BRL-3A增殖。

Abstract

Objective To explore the effect of C7orf42 on cell proliferation of rat hepatocyte line BRL-3A in vitro. Methods The expression of C7orf42 was knocked down by siRNA, and MTT and EdU assay were used to discover the effect of C7orf42 on cell proliferation at 48 hours after transfection. Flow cytometry was used to observe the effect of cell cycle progression. Real-tme PCR and Western blotting were used to detect the changes in the expression of cell proliferation-associated gene. Results MTT results showed that the cell viability of the interference group (C7BRL-siR3) was significantly lower than that of the negative control group (NC) at 48 hours after transfection (P<0.05). Meanwhile, the percentage of EdUp-labeling cells was also significantly decreased (P<0.05). At the same time, the flow cytometry results showed that the number of cells in division phase (S+G₂/M) of the interference group was significantly reduced in parallel (P<0.05). Further, the interference group down-regulated the expression levels of cell proliferation-related genes and proteins of JUN, MYC, CCND1 and CCNA2. Conclusion C7orf42 may promote cell proliferation via regulating the expression of JUN, MYC, CCND1 and CCNA2 in rat hepatocyte line BRL-3A.

关键词

新蛋白 / C7orf42 / 肝细胞系BRL-3A / siRNA / 细胞增殖 / 免疫印迹法 / 大鼠

Key words

Novel protein / C7orf42 / Hepatocyte line BRL-3A / Small interfering RNA / Cell proliferation / Western blotting / Rat

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张春艳常翠芳张世馥马纪张富春徐存拴. 新蛋白C7orf42促进大鼠肝细胞BRL-3A增殖[J]. 解剖学报. 2017, 48(2): 150-155 https://doi.org/10.16098/j.issn.0529-1356.2017.02.005

ZHANG Chun-yan CHANG Cui-fang ZHANG Shi-fu MA Ji ZHANG Fu-chun XU Cun-shuan. Novel protein C7orf42 promotes rat hepatocyte BRL-3A proliferation[J]. Acta Anatomica Sinica. 2017, 48(2): 150-155 https://doi.org/10.16098/j.issn.0529-1356.2017.02.005

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基金

国家自然科学基金;河南省自然科学基金

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