过表达配对相关同源框1(PRRX1)通过调控Wnt/β-catenin通路抑制肝癌细胞体内致瘤性  

Overexpression of paired related homoeobox 1(PRRX1)inhibits tumorigenicity of hepatoma cells in vitro by regulating Wnt/β-catenin pathway

作　　者：尹润龙 尹东亮[1] 卢沛林 李柱威 林志强[1] YIN Run-Long;YIN Dong-Liang;LU Pei-Lin;LI Zhu-Wei;LIN Zhi-Qiang(Department of Hepatobiliary Surgery,Dongguan People′s Hospital,Dongguan 523059,China)

机构地区：[1]东莞市人民医院肝胆外科,东莞523059

出　　处：《中国免疫学杂志》2020年第6期687-692,共6页Chinese Journal of Immunology

基　　金：东莞市社会科技发展(一般)项目(2018507150011277)。

摘　　要：目的:研究慢病毒介导的配对相关同源框1(PRRX1)过表达对肝癌细胞体内致瘤性的影响及相关机制。方法:构建PRRX1过表达的慢病毒载体pLV-PRRX1-IRES2-EGFP,转染HEK293T细胞,获得慢病毒颗粒后,感染肝癌细胞株SMMC-7721,采用Western blot检测感染后细胞株中PRRX1蛋白表达。皮下注射SMMC-7721细胞(对照组、空载体组、PRRX1过表达组及PRRX1过表达+Wnt/β-catenin通路抑制剂XAV939干预组)构建裸鼠肝癌移植瘤模型,其中干预组使用XAV939处理,观察处理后移植瘤体积大小并绘制肿瘤生长曲线;采用HE染色观察各组移植瘤组织病变情况;采用原位末端转移酶标记技术(TUNEL)检测移植瘤中细胞凋亡情况;采用免疫组化检测移植瘤中细胞增殖相关抗原Ki-67的表达;采用Western blot检测移植瘤中PRRX1以及Wnt/β-catenin通路相关蛋白β-catenin、c-Myc的表达。结果:成功构建PRRX1过表达的慢病毒载体并成功感染肝癌细胞株SMMC-7721,感染后细胞中PRRX1蛋白水平明显升高;与空载体组比较,PRRX1过表达组裸鼠移植瘤生长缓慢,组织坏死加重,细胞凋亡及PRRX1蛋白表达增加,而Ki-67、β-catenin及c-Myc蛋白表达均受到抑制;与PRRX1过表达组比较,XAV939干预进一步促进PRRX1过表达对裸鼠移植瘤的作用效果,但不改变PRRX1的表达。结论:过表达PRRX1能显著降低肝癌细胞体内致瘤能力,其机制可能与抑制Wnt/β-catenin信号通路有关。Objective:To investigate the effect of lentivirus mediated paired related homoeobox 1(PRRX1)overexpression on tumorigenicity of hepatoma cells in vitro and its related mechanism.Methods:PRRX1 overexpressed lentivirus vector pLV-PRRX1-IRES2-EGFP was constructed and transfected into HEK293T cells.Lentivirus particles were obtained and infected with hepatoma cell line SMMC-7721.The expression of PRRX1 protein in the infected cell line was detected by Western blot.Subcutaneous injection of SMMC-7721 cells(control group,empty vector group,PRRX1 overexpression group and PRRX1 overexpression+Wnt/β-catenin pathway inhibitor XAV939 intervention group)was used to construct the hepatoma xenograft model of nude mouse.XAV939 treatment was used in the intervention group,and the size of the xenograft volume after treatment was observed and the tumor growth curve was drawn.HE staining was used to observe the pathological changes of xenografts in each group.Apoptosis in xenografts was examined by TUNEL assay.Immuno-histochemistry was used to detect the expression of proliferation-related antigen Ki-67 in xenografts.Western blot was used to assessed the expression of PRRX1 and Wnt/β-catenin pathway related proteinsβ-catenin and c-Myc in xenografts.Results:The lentiviral vector of PRRX1 overexpression was successfully constructed and the hepatoma cell line SMMC-7721 was successfully infected.PRRX1 protein level was significantly increased after infection.Compared with the empty vector group,the growth of xenografts in nude mice in the PRRX1 overexpression group was slow,tissue necrosis was aggravated,apoptosis and PRRX1 protein expression were increased,while the levels of Ki-67,β-catenin and c-Myc protein were inhibited.Compared with PRRX1 overexpression group,XAV939 intervention further promoted the effect of PRRX1 overexpression on xenografts of nude mouse,but did not change the expression of PRRX1.Conclusion:Overexpression of PRRX1 can significantly reduce the tumorigenic ability of hepatoma cells in vitro,and the mechanism

关 键 词：配对相关同源框1 慢病毒载体 肝癌 致瘤性 WNT/Β-CATENIN信号通路 

分 类 号：R735.7[医药卫生—肿瘤]