基础医学与临床 ›› 2016, Vol. 36 ›› Issue (10): 1393-1399.

• 研究论文 • 上一篇    下一篇

乙肝病毒促进肝癌细胞系的转移侵袭力

刘辉,王凤梅,骆莹,王鹏,翟道宽,石文霞,李成龙,朱争艳   

  1. 天津市第三中心医院肝胆疾病研究所,天津市人工细胞重点实验室
  • 收稿日期:2016-02-19 修回日期:2016-04-01 出版日期:2016-10-05 发布日期:2016-09-27
  • 通讯作者: 朱争艳 E-mail:Zhuzhengyan59@163.com
  • 基金资助:
    天津市科委面上项目;天津市科学基金联合资助项目青年项目;天津市卫生局科学基金面上基金;天津市高新技术产业化专项资金

HBV promotes migration and invasion of hepatocellular carcinoma cell lines

  • Received:2016-02-19 Revised:2016-04-01 Online:2016-10-05 Published:2016-09-27
  • Supported by:
    General Program of Tianjin Municipal Science and Technology Commission

摘要: 目的 探讨乙肝病毒(HBV)对肝癌细胞转移能力的影响及其可能机制。方法 以初始汇合度为30%,将3种细胞系HL-7702(人正常肝细胞系)、HepG2(未转染HBV-DNA的人肝癌细胞系)、HepG2.2.15(稳定转染HBV-DNA的人肝癌细胞系)种植于96孔板中,待细胞增殖至70%汇合时,利用划痕器制造划痕伤口,置于活细胞动态成像系统中进行多时间点的显微拍照与数据采集,计算相对伤口密度(RWD),并通过免疫荧光染色与Western blot技术测定细胞中EphA2蛋白表达,分析其与RWD值的相关性。结果 细胞迁移实验中,划痕后24~96 h,HL-7702组RWD显著高于HepG2与HepG2.2.15组(P<0.01),划痕后72~144 h,HepG2.2.15组RWD显著高于HepG2组(P<0.01);细胞侵袭实验中,HL-7702细胞因不能穿过基质胶,而无RWD值;划痕后72~144 h,HepG2.2.15组RWD显著高于HepG2组(P<0.05或P<0.01)。EphA2表达:与HL-7702组比较,HepG2与HepG2.2.15组细胞中EphA2表达水平显著升高(P<0.01),其中HepG2.2.15组中EphA2表达水平显著高于HepG2组(P<0.01),且两组肝癌细胞中EphA2的表达量与划痕实验的RWD值呈显著正相关(迁移实验:P<0.01;侵袭实验:P<0.01)。结论 乙肝病毒可能促进肝癌细胞的迁移和侵袭能力,其机制可能与上调EphA2的异常表达有关。

关键词: 肝癌, 乙肝病毒, 迁移, 侵袭, EphA2

Abstract: Objective To observe and compare the different migration and invasion activities between the HepG2 and HepG2.2.15 cells and to explore the influence and mechanism of HBV on the migration and invasion of hepatocellular carcinoma cells via the live cell imaging technology. Methods The three cell lines of HL-7702, HepG2 and HepG2.2.15 were planted on 96-well plates with a 30% degree of cell confluence. When the cells were in the 70% degree of confluence, the scratch-wound healing assay was performed to determine the relative wound density (RWD) via the live cell imaging technology. The EphA2 expression was examined via immunofluorescence staining and Western blotting and the relationship between the EphA2 expressions and the RWD levels in the HepG2 and HepG2.2.15 groups were analyzed. Results Cell migration assays showed that at 24~96 h after scratch, the RWD in the HL-7702 group was higher than that in the HepG2 and HepG2.2.15 groups (P<0.01). At 72~144 h after scratch, the RWD in the HepG2.2.15 group was higher than that in the HepG2 group (P<0.01). Cell invasion assays showed that there was no quantitative value of RWD in the HL-7702 group. At 72~144 h after scratch, the RWD in the HepG2.2.15 group was higher than that in the HepG2 group (P<0.05 or P<0.01). EphA2 expressions showed that comparing with that in the HL-7702 group, EphA2 expressions increased in the HepG2 and HepG2.2.15 groups (P<0.01), and the EphA2 levels in the HepG2.2.15 group were higher than that in the HepG2 group (P<0.01). Moreover, the EphA2 expression in the HepG2 and HepG2.2.15 groups was positively correlated with the RWD levels in cell migration assays (r=0.962, P=0.002) and cell invasion assays (r=0.980, P=0.001). Conclusions HBV might promote the migration and invasion of hepatocellular carcinoma cells and its mechanism might be associated with the up-regulation of EphA2 expressions.

Key words: Hepatocellular Carcinoma, Hepatitis B Virus (HBV), Migration, Invasion, EphA2

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