配对相关同源框1蛋白在肝癌中的表达及其过表达对肝癌细胞侵袭转移的影响

Expression of paired related homoeobox1 in hepatocellular carcinoma and the effect of its overexpression on the invasion and metastasis of hepatocellular carcinoma cells

目的探讨配对相关同源框1(paired related homoeobox 1,PRRX1)蛋白在肝细胞癌中表达的临床意义以及其过表达对肝癌SMMC7721细胞侵袭转移的影响。方法采用免疫组化检测65例肝细胞癌患者癌组织及配对癌旁组织中PRRX1的表达,并分析其表达与肝细胞癌临床病理参数及生存曲线的关系。通过慢病毒介导PRRX1过表达载体感染肝癌SMMC7721细胞,应用qRT-PCR和Western blot技术检测慢病毒感染后细胞中PRRX1 mRNA和蛋白的表达变化,采用CCK-8法检测细胞活性,Transwell实验检测细胞侵袭和迁移能力,Western blot实验检测细胞中MMP-2、MMP-9以及上皮间质转化标志蛋白E-cadherin和Vimentin的表达。结果PRRX1在肝癌组织中表达显著低于癌旁组织(P<0.05),其表达水平与肝癌患者血管侵犯、肝内转移、远端转移和TNM分期等临床病理特征参数有相关性(P<0.05)。PRRX1阴性表达肝癌患者5年生存率显著低于PRRX1阳性表达肝癌患者(P<0.05)。慢病毒介导PRRX1过表达载体感染可显著增强SMMC7721细胞中PRRX1蛋白和mRNA表达水平(P<0.05),降低SMMC7721的细胞活性以及侵袭和迁移能力,并可显著下调MMP-2、MMP-9、Vimentin蛋白表达,同时显著上调E-cadherin蛋白表达。结论PRRX1在肝癌组织中低表达,其表达水平与肝癌患者血管侵犯、肝内转移、远端转移及TNM分期相关,过表达PRRX1可抑制肝癌SMMC7721细胞侵袭转移。

Objective To investigate the clinical significance of paired related homoeobox1 expression in hepatocellular carcinoma(HCC)and the effect of its overexpression on the invasion and metastasis of HCC cell line SMMC7721.Methods The expression of PRRX1 in 65 cases of HCC and matched adjacent tissues were detected by immunohistochemistry,and the relationship between PRRX1 expression and clinicopathological parameters,survival curve of HCC were analyzed.Lentivirus-mediated PRRX1 overexpression vector was infected into SMMC7721 cells,and the expression of PRRX1 mRNA and protein in infected cells was detected by qRT-PCR and Western blot.The cells viability was detected by CCK-8 assay.The invasion and migration ability of cells were detected by Transwell assay.The proteins expression of MMP-2,MMP-9,E-cadherin and Vimentin were evaluated by western blot.Results The expression of PRRX1 in tumor tissue was significantly lower than that in adjacent tissues(P<0.05).The expression level of PRRX1 was significantly correlated with clinicopathological parameters such as vascular invasion,intrahepatic metastasis,distant metastasis and TNM stage(P<0.05).The 5-year survival rate of PRRX1-negative expression patients with HCC was significantly lower than that in the PRRX1-positive expression patients(P<0.05).Lentiviral-mediated PRRX1 overexpression significantly enhanced the protein and mRNA expression of PRRX1 in SMMC7721 cells(P<0.05).Overexpression of PRRX1 suppressed cell viability,invasion and migration in SMMC7721 cells.In addition,overexpression of PRRX1 resulted in decreased the proteins expression of MMP-2,MMP-9 and Vimentin,and induced E-cadherin protein expression.Conclusions PRRX1 is down-regulated in HCC tissues,and its expression level is associated with vascular invasion,intrahepatic metastasis,distant metastasis and TNM stage.Furthermore,the overexpression of PRRX1 can inhibit the invasion and metastasis of SMMC7721 cells.