乙肝病毒促进肝癌细胞系的转移侵袭力

发布时间：2018-05-18 22:45

本文选题：肝癌 + 乙肝病毒　；参考：《基础医学与临床》2016年10期

【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(P0.01),划痕后72~144 h,HepG2.2.15组RWD显著高于HepG2组(P0.01);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(P0.05或P0.01)。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(P0.01),其中HepG2.2.15组中Eph A2表达水平显著高于HepG2组(P0.01),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:P0.01;侵袭实验:P0.01)。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。
[Abstract]:Objective to investigate the effect of hepatitis B virus (HBV) on metastasis of hepatoma cells and its possible mechanism. Methods three cell lines HL-7702 (human normal liver cell line HL-7702 (human hepatoma cell line without HBV-DNA), HepG2.2.15 (human hepatoma cell line stably transfected with HBV-DNA) were seeded into 96-well plate with an initial confluence degree of 30, and the cells proliferated to 70% confluence. The scratch wound was made by scratch device. The micrograph and data acquisition of multiple time points were carried out in the living cell dynamic imaging system. The relative wound density was calculated and the expression of Eph A2 protein in the cells was determined by immunofluorescence staining and Western blot technique. The correlation between RWD value and it was analyzed. Results in the cell migration test, the RWD of HL-7702 group was significantly higher than that of HepG2 and HepG2.2.15 group (P 0.01), and the RWD of 72144h HepG2.2.15 group was significantly higher than that of HepG2 group (P 0.01) in 24 h after scratch, the HL-7702 cells had no RWD value because they could not penetrate the matrix gel. The expression of Eph A2 in HepG2.2.15 group was significantly higher than that in HepG2 group (P0.05 or P0.01).Eph A2). Compared with HL-7702 group, the expression level of Eph A2 in HepG2 and HepG2.2.15 group was significantly higher than that in HL-7702 group. The expression level of Eph A2 in HepG2.2.15 group was significantly higher than that in HepG2 group, and Eph A2 expression in HCC cells in both groups was significantly higher than that in HL-7702 group. There was a significant positive correlation between the expression of A2 and the RWD value of scratch test (migration experiment: P0.01; invasive experiment: P0.01). Conclusion Hepatitis B virus may promote the migration and invasion of hepatoma cells, and its mechanism may be related to the up-regulation of Eph A2 expression.
【作者单位】：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;
【基金】：天津市科委面上项目(13JCYBJC22500) 天津市科学基金联合资助项目(15JCQNJC45700) 天津市卫生局科学基金(2012WSJ059) 天津市高新技术产业化专项资金[津发改高技(2006)26号]
【分类号】：R735.7;R512.62

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