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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 investigate the effect of unknown function protein C7orf42 on the proliferation of rat hepatocyte BRL-3A cultured in vitro. Methods The expression of C7orf42 was down-regulated by gene interference. The effect of C7orf42 on the proliferation of BRL-3A cells was detected by MTT and Ed U incorporation. The cell cycle progression was detected by flow cytometry. The expression of proliferation related genes was detected by Reat-time PCR and Western blotting. Results MTT assay showed that the viability of cells in interference group was 11% lower than that in negative control group 48 h after transfection with C7orf42 interference fragment (P <0.05). Ed U incorporation assay showed that the ratio of Ed U positive cells in experimental group was significantly higher than that in control group (P <0.05). The results of flow cytometry showed that the number of cells in S + G2 / M phase in the intervention group decreased by 12% (P <0.05). The results of Reat-time PCR showed that the interference of C7orf42 The expressions of JUN, CCND1, MYC and CCNA2 were down-regulated by 26%, 31%, 37% and 14% respectively (P <0.05). The results of Western blotting showed that the expressions of JUN, CCND1 , And MYC and CCNA2 were down-regulated by 59%, 54%, 18% and 27%, respectively (P <0.05). Conclusion C7orf42 may promote BRL-3A proliferation in rat hepatocytes cultured in vitro by up-regulating the expression of JUN, CCND1, MYC and CCNA2.