摘要
目的探讨缺氧诱导基因2(hypoxia-inducible gene 2,HIG2)在肝癌发生发展中的作用和意义。方法正常肝细胞(QZG)、肝癌细胞系(HepG2、SMMC7721)常规培养,采用Western Blot法检测HIG2蛋白的表达水平;通过甘油三酯检测试剂盒检测每6×106个正常肝细胞系(QZG)和肝癌细胞系(HepG2、SMMC7721)甘油三酯含量;使用涂有Matrigel的聚碳酸酯膜的Transwell小室测定细胞侵袭和迁移能力,划痕实验比较转染了对照质粒以及转染了HIG2过表达质粒的HepG2细胞的细胞迁移率;实时定量荧光PCR分析30例肝癌组织和对应肝癌癌旁组织中HIG2蛋白的表达;免疫组织化学方法检测肝癌癌旁组织和肝癌组织中HIG2蛋白的表达及部位。结果肝癌细胞系HepG2和SMMC7721比正常肝细胞系QZG中HIG2蛋白水平增加(P<0.05),同时肝癌细胞系HepG2和SMMC7721甘油三酯含量[(16.94±0.33)mg/dL,(16.04±1.02)mg/dL]均显著高于正常肝细胞系QZG[(4.90±0.39)mg/dL],差异比较具有统计学意义(P<0.05);与转染对照质粒组比较,HIG2的过表达质粒显著增加了HepG2细胞的迁移率和划痕愈合百分比(P<0.05);实时定量荧光PCR结果显示与肝癌癌旁组织比较,肝癌组织中HIG2的丰度增加(P<0.05);Western Blot结果显示与肝癌癌旁组织比较,肝癌组织中HIG2蛋白水平增加(P<0.05);免疫组化显示HIG2在肝癌组织中高表达且主要在胞浆中。结论HIG2蛋白可以显著诱导肝癌细胞的甘油三酯的堆积,同时促进了肝癌细胞的侵袭和转移潜力。
Objective To investigate the effect and significance of hypoxia-inducible gene 2(HIG2)protein in the development of liver cancer.Methods Normal hepatocytes(QZG)and hepatoma cell lines(HepG2,SMMC7721)were routinely cultured,and the expression level of HIG2 protein was detected by Western Blot.Triglyceride level of each 6×106 QZG,HepG2 and SMMC7721 cell lines were detected by triglyceride detection kit.Cell invasion ability and migration rate were measured using a Transwell chamber containing a Matrigel-coated polycarbonate membrane in the HepG2 cell lines that were transfected with control plasmid or HIG2 overexpression plasmid.The expression of HIG2 in 30 cases of hepatocellular carcinoma(HCC)tissues and corresponding paracancerous tissue of HCC were detected by real-time quantitative fluorescent PCR.The expression of HIG2 was detected by immuno-histochemical staining in HCC tissues.Results The levels of HIG2 protein in hepatoma cell lines HepG2 and SMMC7721 were significantly higher than those in QZG cell lines(P<0.05),and the triglyceride levels in HepG2 and SMMC7721 cell lines[(16.94±0.33)mg/dL,(16.04±1.02)mg/dL]were significantly higher than those in QZG cell lines(4.90±0.39)mg/dL.Compared with control plasmid group,overexpression plasmid of HIG2 significantly increased the migration and scratch healing percentage of HepG2 cells(P<0.05).The abundance of HIG2 gene and the protein level of HIG2 in HCC tissues were increased compared with the adjacent tissues of liver cancer(P<0.05)by Western Blot.HIG2 was highly expressed in HCC tissues and mainly in the cytoplasm by immunohistochemistry.Conclusion HIG2 protein can significantly induce the accumulation of triglycerides in HCC cells,and promote the invasion and metastasis potential of HCC.
作者
刘咏真
康静波
杜锐
郑伟
LIU Yongzhen;KANG Jingbo;DU Rui;ZHENG Wei(Tumor Diagnosis and Treatment Center,the Sixth Medical Center,Chinese PLA General Hospital,Beijing 100048,China)
出处
《转化医学杂志》
2020年第3期137-141,146,共6页
Translational Medicine Journal
基金
国家重点研发计划项目(2017YFC0113700)。
关键词
肝癌
缺氧诱导基因2
甘油三酯
肿瘤标志物
Hepatocellular carcinoma(HCC)
Hypoxia-inducible gene 2(HIG2)
Trigly-ceride
Tumor marker
