新蛋白C7orf42促进大鼠肝细胞BRL-3A增殖

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%(P0.05);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%(P0.05);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%(P0.05);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%(P0.05);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%(P0.05)。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。

Novel protein C7orf42 promotes rat hepatocyte BRL-3A proliferation

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 EdU-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.