發(fā)布時間：2018-05-18 22:45

  本文選題：肝癌 + 乙肝病毒　； 參考：《基礎醫(yī)學與臨床》2016年10期

【摘要】：目的探討乙肝病毒(HBV)對肝癌細胞轉(zhuǎn)移能力的影響及其可能機制。方法以初始匯合度為30%,將3種細胞系HL-7702(人正常肝細胞系)、HepG2(未轉(zhuǎn)染HBV-DNA的人肝癌細胞系)、HepG2.2.15(穩(wěn)定轉(zhuǎn)染HBV-DNA的人肝癌細胞系)種植于96孔板中,待細胞增殖至70%匯合時,利用劃痕器制造劃痕傷口,置于活細胞動態(tài)成像系統(tǒng)中進行多時間點的顯微拍照與數(shù)據(jù)采集,計算相對傷口密度(RWD),并通過免疫熒光染色與Western blot技術測定細胞中Eph A2蛋白表達,分析其與RWD值的相關性。結果細胞遷移實驗中,劃痕后24~96 h,HL-7702組RWD顯著高于HepG2與HepG2.2.15組(P0.01),劃痕后72~144 h,HepG2.2.15組RWD顯著高于HepG2組(P0.01);細胞侵襲實驗中,HL-7702細胞因不能穿過基質(zhì)膠,而無RWD值;劃痕后72~144 h,HepG2.2.15組RWD顯著高于HepG2組(P0.05或P0.01)。Eph A2表達:與HL-7702組比較,HepG2與HepG2.2.15組細胞中Eph A2表達水平顯著升高(P0.01),其中HepG2.2.15組中Eph A2表達水平顯著高于HepG2組(P0.01),且兩組肝癌細胞中Eph A2的表達量與劃痕實驗的RWD值呈顯著正相關(遷移實驗:P0.01;侵襲實驗:P0.01)。結論乙肝病毒可能促進肝癌細胞的遷移和侵襲能力,其機制可能與上調(diào)Eph A2的異常表達有關。
[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.
【作者單位】： 天津市第三中心醫(yī)院肝膽疾病研究所天津市人工細胞重點實驗室;
【基金】：天津市科委面上項目(13JCYBJC22500) 天津市科學基金聯(lián)合資助項目(15JCQNJC45700) 天津市衛(wèi)生局科學基金(2012WSJ059) 天津市高新技術產(chǎn)業(yè)化專項資金[津發(fā)改高技(2006)26號]
【分類號】：R735.7;R512.62

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