T淋巴瘤侵袭转移诱导因子1对肝癌细胞迁移和黏附的影响及其机制  被引量:2

Effect of T-lymphoma invasion and metastasis inducible factor 1 on migration and adhesion of hepatoma cells and its molecular mechanism

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作  者:刘云鹤[1] 蔡金保[2] 张德重[3] 满永宏[4] Liu Yunhe;Cai Jinbao;Zhang Dezhong;Man Yonghong(Department of Oncology,the First Affiliated Hospital of Nanyang Medical College,Nanyang 473000,China;Department of Pharmacy,the First Affiliated Hospital of Nanyang Medical College,Nanyang 473000,China;Department of Gastrointestinal Surgery,the First Affiliated Hospital of Xinxiang Medical College,Xinxiang 453000,China;Scientific Research Experiment Center of Nanyang Medical College in Henan Province,Nanyang 473000,China)

机构地区:[1]河南省南阳医学高等专科学校第一附属医院肿瘤科,473000 [2]河南省南阳医学高等专科学校第一附属医院药学部,473000 [3]新乡医学院第一附属医院胃肠外科,453000 [4]河南省南阳医学高等专科学校科研实验中心,473000

出  处:《中华实验外科杂志》2020年第6期1122-1125,共4页Chinese Journal of Experimental Surgery

摘  要:目的探讨T淋巴瘤侵袭转移诱导因子1(Tiam1)对肝癌细胞迁移和黏附的影响及其机制。方法选取2015年6月到2018年6月南阳医学高等专科学校第一附属医院手术切除的104例肝癌组织和癌旁组织作为研究对象,采用蛋白质印迹法(Western blot)和荧光定量聚合酶链反应(PCR)分析肿瘤组织和癌旁组织中Tiam1蛋白和mRNA表达水平;采用规律间隔成簇短回文重复序列及其相关核酸酶9系统(CRISPR-Cas9)技术在肝癌细胞株HepG2和MHCC-97H中建立Tiam1敲除细胞株,分别命名为HepG2 Tiam1 KO组和HepG2对照组;MHCC-97H Tiam1 KO组和MHCC-97H对照组。采用Transwell分析肝癌细胞迁移能力;采用基质黏附实验分析肝癌细胞的黏附能力;采用Western blot分析迁移和黏附相关蛋白的表达水平。组间比较采用t检验。结果与癌旁组织中Tiam1蛋白和mRNA表达水平(1.49±0.21、1.07±0.19)比较,肝癌组织中Tiam1蛋白和mRNA表达水平(3.10±0.37、2.87±0.31)显著增加,差异有统计学意义(t=3.058、2.891,P<0.05)。与HepG2对照组和MHCC-97H对照组细胞迁移数量[(142.17±20.48)、(169.10±24.19)个]比较,HepG2 Tiam1 KO组和MHCC-97H Tiam1 KO组细胞迁移数量[(76.91±9.87)、(91.55±7.52)个]显著下降,差异有统计学意义(t=4.128、3.944,P<0.05)。与HepG2对照组和MHCC-97H对照组细胞黏附能力(142.17±20.48、169.10±24.19)比较,HepG2 Tiam1 KO组和MHCC-97H Tiam1 KO组细胞黏附能力(76.91±9.87、91.55±7.52)显著下降,差异有统计学意义(t=4.128、3.944,P<0.05)。与HepG2对照组和MHCC-97H对照组细胞黏着斑激酶(FAK)蛋白表达水平(1.09±0.19、1.59±0.24)比较,HepG2 Tiam1 KO组和MHCC-97H Tiam1 KO组细胞FAK蛋白表达水平(0.39±0.11、0.44±0.19)显著下降,差异有统计学意义(t=3.208、3.011,P<0.05)。结论Tiam1通过影响FAK蛋白表达水平,进而调控肝癌细胞的迁移和黏附能力。Objective To observe the effect of T-lymphoma invasion and metastasis inducible factor 1(Tiam1)on the migration and adhesion of hepatoma cells and its molecular mechanism.Methods From June 2015 to June 2018,104 cases of liver cancer and adjacent tissues were selected as the research objects in the First Affiliated Hospital of Nanyang Medical College from June 2015 to June 2018.Western blotting and fluorescence quantitative polymerase chain reaction(PCR)were used to analyze the expression level of Tiam1 protein and mRNA in tumor and adjacent tissues.Tiam1 knockout cell lines in HepG2 and MHCC-97H were establish by regularly spaced clustered short palindrome repeats and their related nuclease 9 system(CRISPR/Cas9)technology and then named HepG2 Tiam1 KO group and HepG2 control group;MHCC-97H Tiam1 KO group and MHCC-97H control group,respectively.The migration ability of liver cancer cells was analyzed by Transwell.The adhesion ability of liver cancer cells was analyzed by matrix adhesion test.The expression level of migration and adhesion related proteins was analyzed by Western blotting.Results Compared with the adjacent tissues(1.49±0.21,1.07±0.19),the expression level of Tiam1 protein and mRNA(3.10±0.37,2.87±0.31)in liver cancer tissues significantly increased(t=3.058,2.891,P<0.05).Compared with HepG2 control group and MHCC-97H control group,the number of migrating cells in HepG2 Tiam1 KO group and MHCC-97H Tiam1 KO group significantly decreased(t=4.128,3.944,P<0.05).Compared with HepG2 control group and MHCC-97H control group,cell adhesion in HepG2 Tiam1 KO group and MHCC-97H Tiam1 KO group significantly decreased(t=4.128,3.944,P<0.05).Compared with the HepG2 and MHCC-97H control groups(1.09±0.19,1.59±0.24),the expression level of focal adhesion kinase(FAK)in HepG2 Tiam1 KO group and MHCC-97H Tiam1 KO group(0.39±0.11,0.44±0.19)significantly decreased(t=3.208,3.011,P<0.05).Conclusion Tiam1 can regulate the migration and adhesion of hepatoma cells by influencing the expression of FAK protein.

关 键 词:T淋巴瘤侵袭转移诱导因子1 肝癌 迁移 黏附 

分 类 号:R735.7[医药卫生—肿瘤]

 

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