RAD51AP1沉默通过调控有丝分裂基因抑制肝癌细胞增殖

RAD51AP1 silencing inhibits the proliferation of hepatocellular carcinoma cells by regulating mitosis genes

目的分析沉默RAD51AP1基因对肝癌细胞增殖的影响,并初步探索其作用机制。方法利用qPCR检测RAD51AP1在不同肝癌细胞系中的表达;制备shRAD51AP1慢病毒并转导HepG2和Huh7细胞来沉默RAD51AP1,采用qPCR检测RAD51AP1的沉默效率;MTS法和克隆形成实验检测RAD51AP1沉默对细胞增殖和克隆形成能力的影响;根据转录组测序结果筛选出差异表达基因,并作进一步的检测分析;通过裸鼠体内成瘤实验研究RAD51AP1沉默对肝癌细胞增殖的影响。结果MTS结果显示,HepG2和Huh7细胞中RAD51AP1沉默组的增殖能力均显著低于对照组(P<0.01);克隆形成实验显示,RAD51AP1沉默组克隆形成数明显少于对照组(P<0.01);通过转录测序筛选出差异表达基因TACC1、NEK7,并从蛋白水平进行了验证。结论RAD51AP1基因沉默能够有效抑制肝癌细胞增殖,有望成为预防或治疗肝癌的新靶点。

Objective To explore the effects of silencing RAD51AP1 gene on the proliferation of hepatocellular carcinoma(HCC)cells,and to initially explore its inhibition mechanism.Methods The qPCR method was used to determine the specific expression level of RAD51AP1 in different HCC cell lines.The shRAD51AP1 lentivirus was prepared and transferred into HepG2 and Huh7 cells to silence the RAD51AP1 gene.The silencing efficiency of RAD51AP1 was detected by qPCR,and the effects of silencing RAD51AP1 on cell proliferation and colony formation was detected by MTS and colony formation assay respectively.The differentially expressed genes were screened out according to the transcriptome sequencing results,and further analyzed.The effects of silencing RAD51AP1 on the proliferation of hepatocarcinoma cells were also studied by the tumorigenesis experiment in nude mice.Results The MTS results showed that the proliferation ability of HepG2 and Huh7 cells in the RAD51AP1 silence group was significantly lower than that in the control group(P<0.01).The colony formation assay results showed that the number of clones was significantly less in the RAD51AP1 silence group than in the control group(P<0.01).Through transcriptome sequencing,the differentially expressed genes TACC1 and NEK7 were screened out,and verified by Western blotting at protein level.Conclusion Silencing RAD51AP1 gene can effectively inhibit the proliferation of HCC cells.It may become a new target for the prevention or treatment of HCC.