SYT1对肝癌细胞侵袭、转移的影响及其分子机制

Effect of SYT1 on Invasion and Metastasis of Liver Cancer Cells and its Molecular Mechanism

目的:研究突触囊泡蛋白Ⅰ(SYT1)对肝癌侵袭转移的影响及其机制。方法:选择人肝癌细胞(Hep3B、Huh7、HCCLM3、SMMC-7721、HLF和BEL-7404)和人正常肝细胞HL-7702作为研究对象,采用实时荧光定量聚合酶链反应实验(q PCR)检测SYT1在这些细胞中的表达;选取150例原发性肝癌组织及其对应癌旁组织作为研究对象,通过免疫组化实验分析SYT1在这两种组织中的表达;选择肝癌细胞Hep3B、Huh7作为观察对象,根据转染的载体不同分别将其分为对照组(转染表达空载体)、干扰组(转染干扰质粒)和过表达组(转染过表达质粒),通过Transwell迁移实验和OrisTM细胞迁移实验检测SYT1对肝癌细胞迁移能力的影响,通过Transwell侵袭实验检测SYT1对肝癌细胞侵袭能力的影响,采用Western blot实验检测上皮-间充质细胞转化(EMT)相关蛋白钙黏附蛋白E(E-cadherin)和波形蛋白(Vimentin)的表达。结果:SYT1的m RNA和蛋白在肝癌组织中的表达高于对应的癌旁组织(P<0.05);从基因水平检测SYT1的表达,SYT1在肝癌细胞中的表达高于正常肝细胞,Huh7和Hep3B细胞中SYT1的表达高于其他肝癌细胞(P<0.05);与对照组比较,干扰组的肝癌细胞迁移、侵袭能力降低,过表达组的肝癌细胞迁移、侵袭能力增强(P<0.05);与对照组比较,干扰组的肝癌细胞E-cadherin蛋白表达增多,Vimentin蛋白表达量减少(P<0.05)。结论:干扰SYT1的表达能抑制肝癌细胞EMT过程,进而抑制其侵袭、迁移能力。

Objective:To investigate the effect of synaptotagmin I(SYT1)on the invasion and metastasis of liver cancer cells and its mechanism.Methods:Hepatocellular carcinoma cells(Hep3 B,Huh7,HCCLM3,SMMC-7721,HLF,and BEL-7404)and human normal liver cells HL-7702 were selected as the research objects,real-time quantitative polymerase chain reaction(qPCR)was used to detect the expression of SYT1 in these cells.150 cases of primary liver cancer tissues and their corresponding adjacent tissues were selected as the research objects.The differential expression of SYT1 was analyzed by immunohistochemistry.Hep3 B and Huh7 cells were selected as the observation objects.According to the different vectors,they were divided into control group(empty vector),interference group(interference plasmid)and overexpression group(overexpression plasmid),the effects of SYT1 on the invasion and migration ability of liver cancer cells were observed by Transwell and OrisTMcell migration experiments after interfering with the expression of SYT1.Transwell invasion assay was used to detect the effect of SYT1 on the invasion of HCC cells.Results:The expression of SYT1 m RNA and protein in hepatocellular carcinoma was higher than that in adjacent tissues(P<0.05).The expression of SYT1 was detected by qPCR,the expression of SYT1 in hepatocellular carcinoma cells was higher than that in normal liver cells,and the expression of SYT1 in Huh7 and Hep3 B cells was higher than that in other liver cancer cells(P<0.05).Compared with the control group,the migration and invasion ability of HCC cells in the interference group was decreased,and the migration and invasion ability of HCC cells in the overexpression group was enhanced,the differences were statistically significant(P<0.05);compared with the control group,the expression of Ecadherin protein in the interference group was increased,and the expression of vimentin protein was decreased,the differences were statistically significant(P<0.05).Conclusion:Interference with the expression of SYT1 can inhibit the epithelial-mesenchymal cell transformation process of liver cancer cells,thereby inhibiting their invasion and metastasis.