新蛋白C7orf42促进大鼠肝细胞BRL-3A增殖  

Novel protein C7orf42 promotes rat hepatocyte BRL-3A proliferation

作　　者：张春艳[1,2] 常翠芳[2] 张世馥[2] 马纪[1] 张富春[1] 徐存拴 

机构地区：[1]新疆大学生命科学与技术学院,新疆生物资源基因工程重点实验室,乌鲁木齐830046 [2]河南师范大学省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

出　　处：《解剖学报》2017年第2期150-155,共6页Acta Anatomica Sinica

基　　金：国家自然科学基金(31572270);河南省自然科学基金(142300413227)

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、Ed U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);Ed U掺入法检测表明,实验组的Ed U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。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 Ed U 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 Ed U-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 

分 类 号：Q28[生物学—细胞生物学]