C-X-C基序趋化因子10表达与肝癌患者预后的关系及其下调对肝癌增殖和侵袭的影响  

Association of C-X-C motif chemokine 10 expression with the prognosis of patients with hepatocellular carcinoma and the effects of C-X-C motif chemokine 10 downregulation on cell proliferation and invasion

作　　者：林锐[1] 温培豪[1] 李晓丽[1] Lin Rui;Wen Peihao;Li Xiaoli(Department of General Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院普外科,450052

出　　处：《中华实验外科杂志》2021年第2期332-335,共4页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关计划省部共建项目 (SB201901014);河南省医学科技攻关计划联合共建项目 (LHGJ20190068)。

摘　　要：目的检测C-X-C基序趋化因子10(CXCL10)在人肝癌组织中的表达水平及其与肝癌患者临床生物学特征和预后的关系,并探讨CXCL10对肝癌细胞增殖和侵袭的作用。方法癌症基因组图谱(TCGA)RNA测序数据库分析CXCL10在配对(n=56)、非配对肝癌组织(n=370)、癌旁正常组织(n=56)的表达水平,根据CXCL10表达水平、生存时间和生存状态获得CXCL10在肝癌的临界值,根据其cutoff值我们将肝癌患者分为CXCL10高表达组和CXCL10低表达组,进一步分析其与肝癌患者临床生物学特征、生存和肿瘤复发的关系。利用小干扰RNA在肝癌HepG2细胞株敲低CXCL10基因,实时定量聚合酶链反应(PCR)和蛋白质印迹法(Western blot)检测CXCL10、增殖细胞核抗原(PCNA)和基质金属蛋白酶(MMP)-2的表达水平。噻唑蓝(MTT)和Transwell检测CXCL10沉默对肝癌细胞增殖和侵袭的影响。两组间比较采用独立t检验。χ^(2)检验分析CXCL10表达与肝癌临床病理学特征的关系;Kaplan-Meier及Cox回归分析CXCL10表达与肝癌患者预后的关系。结果t检验分析结果显示,CXCL10在配对肝癌组织(t=3.677,P<0.01)和非配对肝癌组织的表达水平显著高于癌旁组织(t=4.692,P<0.01);χ^(2)检验显示,其高表达与肝癌患者年龄(χ^(2)=4.680,P<0.05)、性别(χ^(2)=7.993,P<0.01)、肿瘤大小(χ^(2)=7.436,P<0.05)及病理学分级(χ^(2)=6.350,P<0.05)均明显相关,但与其他临床病理学因素无相关(χ^(2)=0.092、0.468,P>0.05);Kaplan-Meier分析显示,CXCL10高表达患者的生存率低于CXCL10低表达患者(χ^(2)=4.672,P<0.05)而肿瘤复发率高于CXCL10低表达患者(χ^(2)=4.672,P<0.05),差异有统计学意义。体外实验显示,CXCL10沉默减少PCNA和MMP-2蛋白表达,抑制肝癌细胞增殖活性。CXCL10沉默组肝癌侵袭细胞数[(51.2±5.4)个]明显低于空载组[(136.3±11.5)个]穿过小室的细胞数(t=2.315,P<0.01),差异有统计学意义。结论CXCL10高表达与肝癌患者年龄、性别、肿瘤大小、�Objective To examine the expression levels of C-X-C motif chemokine 10(CXCL10)in hepatocellular carcinoma(HCC)tissues,analyze its association with the clinicopathological parameters and prognosis in patients with HCC,and assess the effects of CXCL10 downregulation on cell proliferation and invasion.Methods The expression levels of CXCL10 in paired and unpaired HCC tissues and adjacent normal tissues were examined by the cancer genome atlas(TCGA)RNA-seq data.HCC cell line HepG2 stably transfected with CXCL10 small interfering RNA(siRNA)(siCXCL10)was constructed and the expression levels of CXCL10,proliferating cell nuclear antigen(PCNA)and metalloproteinase(MMP)-2 were detected by quantitative Real-time PCR(RT-qPCR)and Western blotting assays.The proliferative activity and metastatic capability of HepG2 cells were evaluated by methyl thiazolyl tetrazolium(MTT)and Transwell assays.Results T test showed that the expression levels of CXCL10 were increased in paired(t=3.677,P<0.01)and unpaired(t=4.692,P<0.01)HCC tissues,as compared with the adjacent normal tissues.χ^(2) analysis showed that high expression of CXCL10 was associated with the age(χ^(2)=4.680,P<0.05),gender(χ^(2)=7.993,P<0.01),tumor size(χ^(2)=7.436,P<0.05)and pathological stage(χ^(2)=6.350,P<0.05),but had no association with other clinicopathological factors in HCC patients(χ^(2)=0.092,0.468,P>0.05).Kaplan-Meier analysis showed that the patients with high CXCL10 expression had a lower survival rate(χ^(2)=4.672,P<0.05)but a higher tumor recurrence rate(χ^(2)=4.672,P<0.05)than in those with low CXCL10 expression.Silencing of CXCL10 gene downregulated the expression of PCNA and MMP-2,and inhibited the proliferative viability and invasive potential of HepG2 cells(t=2.315,P<0.01).Conclusion High expression of CXCL10 is associated with the age,gender,tumor size,pathological stage and poor prognosis in patients with HCC,and its knockdown suppresses the proliferation and invasion of HCC cells.

关 键 词：肝癌 预后 增殖 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]