乙肝病毒促进肝癌细胞系的转移侵袭力

HBV promotes migration and invasion of hepatocellular carcinoma cell lines

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Eph A2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）。Eph A2表达：与HL-7702组比较,HepG2与HepG2.2.15组细胞中Eph A2表达水平显著升高（P〈0.01）,其中HepG2.2.15组中Eph A2表达水平显著高于HepG2组（P〈0.01）,且两组肝癌细胞中Eph A2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Eph A2的异常表达有关。

Objective To observe and compare the different migration and invasion activities between Hep G2 and Hep G2. 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,Hep G2 and Hep G2. 2. 15 were planted on 96-well plates. When the cells reached 70% confluence,the scratch-wound healing assay was performed to determine the relative wound density（ RWD） via the live cell imaging technology. The Eph A2 expression was examined via immunofluorescence staining and Western blot and the relationship between the Eph A2 expressions and the RWD levels in the Hep G2 and Hep G2. 2. 15 groups were analyzed. Results 24- 96 hafter scratch,the RWD in the HL-7702 group was higher than that in the Hep G2 and Hep G2. 2. 15 groups（ P〈 0. 01）. At 72- 144 h after scratch,the RWD in the Hep G2. 2. 15 group was higher than that in the Hep G2 group（ P〈 0. 01）. There was no quantitative value of RWD in the HL-7702 group. At 72- 144 h after scratch,the RWD in the Hep G2. 2. 15 group was higher than that in the Hep G2 group（ P 0. 05 or P〈 0. 01）. Eph A2 expressions showed that comparing with that in the HL-7702 group,Eph A2 expressions increased in the Hep G2 and Hep G2. 2. 15 groups（ P〈 0. 01）,and the Eph A2 level in the Hep G2. 2. 15 group was higher than that in the Hep G2 group（ P〈 0. 01）. Moreover,Eph A2 expression in the Hep G2 and Hep G2. 2. 15 groups was positively correlated with the RWD level in cell migration assays（ r = 0. 962,P〈 0. 01） and cell invasion assays（ r = 0. 980,P〈 0. 01）. Conclusions HBV may promote the migration and invasion of hepatocellular carcinoma cells and its mechanism is potentially associated with the up-regulation of Eph A2 expression.