类泛素化蛋白酶1在肝细胞肝癌组织的表达及其与侵袭和转移的关系  

Overexpressions of sentrin -specific protease -1 is associated with the invasion and metastasis of human hepatocellular carcinoma

作　　者：刘延[1] 王国良[1] 栅产 刘江伟[1] 赵鹏伟[1] 李湘竑[2] 杨定华[2] 黄建钊[1] Liu Yan;Wang G uoliang;Liu Yah;Liu Jiangwei;Zhao Pengwei;Li Xianghong;Yang Dinghua;Huang Jianzhao(Department of Hepatobiliary Surgery,Guizhou Provincial People＇s Hospital,Guiyang 550002,China（Liu Y,Wang GL,Liu Y,Liu JW,Zhao PW,Huang JZ;Department of Hepatobiliary Surgery,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China（Li XH,Yang D)

机构地区：[1]贵州省人民医院肝胆外科,贵阳550002 [2]南方医科大学南方医院肝胆外科,广州510515

出　　处：《中华实验外科杂志》2018年第10期1859-1861,共3页Chinese Journal of Experimental Surgery

基　　金：贵州省科技厅基金（黔科合J字[2015]2087号）;贵州省科技创新人才团队项目（黔科合人才团队[201414008号）

摘　　要：目的探讨类泛素化蛋白酶1（SENP-1）在人肝细胞肝癌中的表达及其与肝癌细胞侵袭转移的关系。方法应用聚合酶链式反应（PCR）和免疫组织化学方法检测肝癌组织，癌边缘组织，非癌组织中SENP-1和缺氧诱导因子-1d（HIF-la）的表达，同时检测微血管密度（MVD），使用PCR和Westernblot检测3种肝癌细胞株MHCC97H、MHCC97L和HepG2的SENP-1表达，SENPl-siRNA质粒转染基因沉默高表达SENP-1基因的肝癌细胞MHCC97H，划痕和侵袭试验检测转染前后肝癌细胞迁移和侵袭能力的变化，Westernblot检测转染前后半胱氨酰天冬氨酸特异性蛋白酶（Caspase-8）、B淋巴细胞瘤2基因（bcl-2）和相关下游基因HIF-la和血管内皮生长因子（VEGF）表达变化。结果肝癌边缘组织中SENP-1、HIF-1仅的表达显著高于肝癌组织及非癌组织（P=0．000），癌边缘组织中MVD低于癌组织（P=0．000）。3种肝癌细胞株SENP-1表达均比人永生化肝细胞株高，SENP-1的表达随着肝癌细胞株侵袭转移能力的增加而增高。SENPl-siRNA质粒转染MHCC97H细胞后，促细胞凋亡蛋白Caspase-8表达升高，抑制细胞凋亡蛋白bcl-2表达降低，相关下游基因HIF-la和VEGF的表达降低。划痕试验结果显示，转染后肝癌细胞划痕距离在24h后较未处理组增加。Transwell试验结果显示，转染后肝癌细胞穿膜细胞数较未处理组明显减少（P=0．000）。结论SENP-1在人肝癌细胞肝癌中的高表达具有促进肝细胞癌侵袭转移的作用，其机制可能与SENP-1的过表达促进肝癌细胞适应缺氧微环境和抑制细胞凋亡有关。Objective To investigate the expressions of sentrin specific protease - 1 （ SENP - 1 ） in hepatocellular carcinoma （ HCC）, and explore its correlations with tumor metastasis. Methods The expression of SENP - 1 and hypoxia inducible factor - 1a （ HIF-la） were assessed by polymerase chain reaction （PCR）and immunohistochemical staining in human hepatocellular carcinoma, edge tissues and noncancer tissues, microvascular density （MVD）was determined. The expressions of SENP- 1 in HCC cell lines MHCC97H, MHCC97L and HepG2 were assessed by reverse transcriptase- PCR （RT- PCR）, quantitative real -time quantitative PCR （Real- time PCR） and Western blotting. The plasmid SENP1 - small interfering RNA （siRNA） conducted was used to transfected into MHCC97H cells. Western blotting analysis was identified for the effect of transfection. The expression of cysteinyl aspartate specific proteinase -8 （ Caspase - 8 ）, B - cell lymphoma - 2 （ bcl - 2 ）, HIF-la and vascular endothelial growth factor （VEGF） protein in hepatoma cells before and after transfection was detected by Western blotting. The invasion and metastasis of hepatoma cells were detected by wound assay and Transwell invasion assay. Results Compared with the hepatocellular carcinoma tissues and non - cancer tissues, the expression of SENP - 1 and HIF-la in tumor - surrounding tissues were much higher. Compared with immortalized liver cell line, the HCC cell lines showed higher expression of SENP1 mRNA and protein, the expressions ofSENP- 1 was positive correlation with the metastasis ability of HCC cell lines. After transfection of SENP1 - siRNA plasmid into MHCC97H cells, the expression of apoptosis promoting protein Caspase - 8 was increased, and the expression of apoptotic protein bcl - 2 was inhibited. Scratch healing assay revealed that the scuffing distance of hepatoma cells after transfection for 24 h was significantly increased compared with non - treatment group. Transwell assay showed that the number of hepatoma cells

关 键 词：类泛素化蛋白酶1 肝细胞癌 侵袭转移 

分 类 号：R735.7[医药卫生—肿瘤]