靶向沉默CXCR3对肝癌细胞恶性增殖的作用研究

Study on the effects of target-silencing CXCR3 expression on malignant proliferation of hepatocellular carcinoma

目的明确CXCR3在肝癌组织中的表达及其与患者总体生存情况的关系，并探讨靶向沉默CXCR3基因对肝癌细胞增殖的影响及其作用机制。方法采用免疫组织化学方法检测60例肝癌组织及相应癌旁组织中CXCR3表达情况，分析CXCR3在肝癌组织中的表达水平与肝癌患者临床病理学之间的相关性，并结合随访资料进行单因素Kaplan-Meier生存分析；采用慢病毒LVCXCR3-shRNA病毒感染Huh7肝癌细胞，通过细胞计数试剂盒-8增殖实验及裸鼠荷瘤实验明确CXCR3缺失对肝癌细胞增殖的影响。计量资料的方差齐性检验用F检验，两两比较，方差齐者用t检验，方差不齐者用校正t检验，各组间率的比较用X^2检验，多组等级资料的比较用Wilcoxon秩和检验。结果CXCR3在肝癌组织中高表达，且CXCR3高表达患者的总体生存率显著低于低表达患者。Huh7肝癌细胞内CXCR3基因被有效沉默后，Huh7细胞体外增殖能力明显受到抑制，裸鼠荷瘤生长速度减慢；且Huh7细胞内JAKSTAT通路活性降低，c-MYC与Bcl-xl蛋白水平下降；此外，CXCR3缺失可有效抑制白细胞介素6介导的JAKSTAT通路激活。结论CXCR3高表达提示患者生存率差；靶向沉默CXCR3基因可抑制肝癌细胞增殖，可能与JAK-STAT通路活性抑制有关。CXCR3有可能成为肝癌治疗的潜在靶点。

Objective To explicit, the expression of chemokine receptor 3 in HCC tissues and its relationship with overall survival of patients, and to explore the effect of targeted silencing CXCR3 gene on proliferation of hepatocellular carcinoma cells and its mechanism of action. Methods The expression of CXCR3 in 60 cases of hepatocellular carcinoma and its adjacent tissues were detected by immunohistochemistry. The clinicopathological correlations between the expression levels of CXCR3 in hepatoma tissues of liver cancer patients were analyzed and univariate Kaplan-Meier survival analysis was performed in combination with follow-up data. Huh7 hepatoma cells were infected with lentivirus LV- CXCR3-shRNA. The effects of CXCR3 deletion on proliferation of hepatoma cells were determined by CCK-8 assay and tumor-bearing nude mice experiment. Results CXCR3 was highly expressed in HCC tissues, and the overall survival rate （OS） of patients with high CXCR3 expression was significantly lower than that of patients with low expression. After the CXCR3 gene was successfully silenced in Huh7 hepatocellular carcinoma cells, the proliferation ability of Huh7 cells was significantly inhibited in vitro, and the tumor growth rate of nude mice was slowed down, and the activity of JAK-STAT pathway in Huh7 cells was decreased, and the levels of c-MYC and Bcl-xl protein were decreased. In addition, deletion of CXCR3 can effectively inhibit IL-6-mediated JAK-STAT pathway activation. Conclusion CXCR3 high expression indicated that the survival rate was poor, and the target silencing of CXCR3 gene could inhibit the proliferation of hepatocellular carcinoma cells and maybe related to inhibition of JAK-STAT pathway activity. CXCR3 may be a potential target for the treatment of hepatocellular carcinoma.