敲低FAM83A对结直肠癌细胞增殖能力的影响及机制研究

Effect and mechanism of FAM83A gene knockdown on proliferation of colorectal cancer cells

目的探索序列相似性83家族成员A(family with sequence similarity 83 member A,FAM83A)在结直肠癌的表达情况,和敲低FAM83A对结直肠癌细胞增殖能力的影响及相关机制。方法用免疫组化检测102例结直肠癌患者组织及其癌旁组织标本中FAM83A的表达情况。将HCT116细胞分为实验组和对照组,实验组细胞转染FAM83A-siRNA质粒,对照组细胞转染MOCK-siRNA质粒,荧光定量PCR检测各组细胞FAM83AmRNA含量,蛋白质印迹法检测各组细胞的FAM83A、PI3K、p-AKT、p-mTOR表达情况,CCK8实验和克隆形成实验检测各组细胞的增殖能力。结果果结直肠癌患者组织FAM83A阳性率为88.23%(90/102),癌旁组织中表达率为10.78%(11/102),FAM83A在结直肠癌组织中的表达率显著高于癌旁组织,差异具有统计学意义(P<0.001)。转染siRNA后,实验组和对照组HCT116细胞中FAM83AmRNA表达量分别为1.23±0.20和0.43±0.12,FAM83A蛋白表达量分别为1.19±0.11和0.23±0.08,PI3K的表达量分别为1.21±0.17、0.28±0.09,p-AKT的表达量分别为1.35±0.23、0.57±0.18,p-mTOR表达量分别为1.48±0.20、1.05±0.14,PI3K、p-AKT、p-mTOR表达均下调(均P<0.05)。实验组和对照组HCT116细胞CCK8实验120h的吸光度分别为1.09±0.22和2.21±0.27实验组和对照组HCT116细胞的克隆形成率分别为21.6%±2.4%和62.7%±4.1%,实验组细胞增殖能力明显下降(P<0.05)。结论FAM83A在结直肠癌组织中表达明显升高,可能与结直肠癌的恶性程度相关;且FAM83A通过PI3K/AKT/mTOR信号通路影响结直肠癌细胞的增殖能力。

Objective To explore the expression of family with sequence similarity 83 member A(FAM83A)in colorectal cancer,and the effect of FAM83A knockdown on the proliferation of colorectal cancer cells and the related mechanism.Methods The expression of FAM83A in the tissues of 102 patients with colorec-tal cancer and its adjacent tissues was detected by immunohistochemistry.HCT116 cells were divided into experi-mental group and control group.The experimental group cells were transfected with FAM83A-siRNA plasmid,and the control group cells were transfected with MOCK-siRNA plasmid.The mRNA content of FAM83A in each group was detected by fluorescence quantitative PCR.The expressions of FAM83A,P13K,p-AKT and p-mTOR in each group were detected by Western blot.CCK8 assay and clonogenesis assay were used to detect cell proliferation.Results The positive rate of FAM83A in colorectal cancer patients was 88.23%(90 cases/102 cases),and the expression rate of FAM83A in paracancer tissues was 10.78%(11 cases/102 cases).The expression rate of Fam83a in colorectal cancer tissues was significantly higher than that in paracancer tissues,with statistical significance(P<0.001).After siRNA transfection,the mRNA expression levels of FAM83A in HCT116 cells of the experimental group and control group were 1.23±0.20 and 0.43±0.12,respectively,and the protein expression levels of FAM83A were 1.19±0.11 and 0.23±0.08,respectively.The expression levels of P13K were 1.21±0.17 and 0.28±0.09,the expression levels of p-AKT were 1.35±0.23 and 0.57±0.18,and the expression levels of p-mTOR were 1.48±0.20 and 1.05±0.14.The expression of P13K,p-Akt and p-mTOR was down-regulated(all P<0.05).The absorbance of HCT116 cells in the experimental group and the control group was 1.09±0.22 and 2.21±0.27,respectively.The cloning rate of HCT116 cells in the experimental group and the control group was 21.6%±2.4%and 62.7%±4.1%,respectively.The proliferation ability of HCT116 cells in the experimental group decreased significantly(P<0.05).Conclusions The expression of FAM83A is significantly increased in colorectal cancer tissues,which may be related to the malignant degree of colorectal cancer.FAM83A affects the proliferation of colorectal cancer cells through the P13K/AKT/mTOR signaling pathway.