NFAT5在肝细胞癌中表达的临床意义及其对IL-6、TNF-α表达的调控作用

Clinical significance of NFAT5 expression in hepatocellular carcinoma and its regulatory effect on the expression of inflammatory related factors IL-6 and TNF-α

目的探索活化T细胞核因子5(nuclear factor of activated T cells 5,NFAT5)在肝细胞癌中表达的临床意义及其对炎症相关因子白细胞介素6(interleukin-6,IL-6)、肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)表达的调控作用。方法用免疫组化检测76例肝癌患者组织及其癌旁组织标本中NFAT5的表达情况。将HuH-7细胞分为实验组和对照组,实验组细胞转染NFAT5-siRNA质粒,对照组细胞转染NC-siRNA质粒,荧光定量PCR检测各组细胞NFAT5 mRNA含量,蛋白质印迹法检测各组细胞的NFAT5、IL-6、TNF-α表达情况,CCK8检测各组细胞的增殖能力。结果肝癌患者组织NFAT5阳性率为81.58%(62/76),癌旁组织中表达率为6.58%(5/76),NFAT5在肝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HuH-7细胞中NFAT5 mRNA表达量分别为1.03±0.19和0.33±0.11,NFAT5蛋白表达量分别为0.94±0.14和0.26±0.07,IL-6表达量分别为1.09±0.16和0.49±0.10,TNF-α表达量分别为1.21±0.13和0.28±0.08,IL-6、TNF-α的表达显著下降(均P<0.05)。实验组和对照组HuH-7细胞CCK8实验120 h的吸光度分别为2.21±0.12和1.12±0.11,实验组细胞增殖能力明显下降(P<0.05)。结论NFAT5在肝细胞肝癌中表达明显升高,可能与肝癌的恶性程度相关;且NFAT5通过调控IL-6、TNF-α的表达影响肝细胞肝癌细胞的增殖能力。

Objective To explore the clinical significance of nuclear factor of activated T cells 5(NFAT5)expression in hepatocellular carcinoma and its regulatory effect on the expression of inflammatory related factors IL-6 and TNF-α.Methods Immunohistochemistry was used to detect the expression of NFAT5 in the tissues of 76 patients with liver cancer and their adjacent tissues.HuH-7 cells were divided into experimental group and control group.The cells in the experimental group were transfected with NFAT5-siRNA plasmid,and the cells in the control group were transfected with NC-siRNA plasmid.The content of NFAT5 mRNA in the cells of each group was detected by fluorescence quantitative PCR,and the content of NFAT5 mRNA in each group was detected by Western blotting.The expression of NFAT5,IL6 and TNF-α in the cells,and the proliferation ability of the cells in each group was detected by CCK8.Results The positive rate of NFAT5 in liver cancer tissues was 81.58%(62/76),and the expression rate in adjacent tissues was 6.58%(5/76).The expression rate of NFAT5 in hepatocellular carcinoma tissues was significantly higher than that in adjacent tissues,and the difference was statistically significant.Academic significance(P<0.001).After transfection of siRNA,the expression levels of NFAT5 mRNA in HuH-7 cells in experimental group and control group were 1.03±0.19 and 0.33±0.11,respectively.The protein expression levels of NFAT5 were 0.94±0.14 and 0.26±0.07,respectively,and the expression levels of IL6 were 1.09±0.16 and 0.49±0.10,respectively.The expression levels of TNF-αwere 1.21±0.13 and 0.28±0.08,respectively,and the expressions of IL6 and TNF-α were significantly decreased(all P<0.05).The absorbance of HuH-7 cells in the experimental group and the control group at 120 h was 2.21±0.12 and 1.12±0.11,respectively,and the cell proliferation ability of the experimental group was significantly decreased(P<0.05).Conclusion The expression of NFAT5 is significantly increased in hepatocellular carcinoma,which may be related to the malignancy of hepatocellular carcinoma.And NFAT5 affects the proliferation of hepatocellular carcinoma cells by regulating the expression of IL6 and TNF-α.