体内外共培养体系中肝癌细胞对肝星状细胞活化的影响

Activation of hepaticstellate cells cocultured with hepatoma carcinoma cells in vitro and in vivo

目的探讨共培养体系中肝癌细胞(MHCC97H)对肝星状细胞(hepatic stellate cell, HSC)活化的影响。方法建立MHCC97H细胞与HSC直接接触式共培养、纤维蛋白原-凝血酶法共培养、条件培养液共培养、Transwell小室法共培养体系及两种细胞联合荷裸鼠动物模型,免疫细胞化学染色法与Western blot法检测α-SMA表达,CCK-8法检测HSC增殖能力,划痕法及小室法检测HSC迁移运动能力。结果各种共培养体系中HSC均呈现活化型状态,直接接触式共培养、条件培养液共培养、Transwell小室法共培养体系及细胞联合荷裸鼠动物模型中HSC的α-SMA蛋白表达明显增强;直接接触式共培养、纤维蛋白原-凝血酶法共培养体系及荷裸鼠动物模型中细胞之间的趋化性增强;肝癌细胞条件培养液与Transwell小室共培养体系中HSC的增殖与迁移运动能力均明显增强。结论在成功构建的多种体内外共培养体系中,肝癌细胞能够促进HSC活化、细胞增殖及迁移运动能力。

Objective To investigate the activation of hepatic stellate cells(HSCs)cocultured with hepatoma carcinoma cells.Methods MHCC97H cells and HSCs were cocultured by cell-cell contact method,fibrinogenthrombin paste technique,conditioned media from MHCC97H(MHCC97H-CM)and Transwell coculture technique,respectively.MHCC97H cells and HSCs were inoculated s.c.into nude mice.The expression ofα-SMA in HSCs was assessed by immunocytochemical staining and Western blotting.Proliferation and migration of HSCs was determined using Cell-Counting Kit-8(CCK-8)and wound healing and Transwell technique,respectively.Results The activation of HSCs was significantly increased in the all cocultured system.The expression ofα-SMA was upregulated by cell-cell contact method,MHCC97H-CM,Transwell coculture technique in vitro and in cancer-bearing mice in vivo.The increased chemotaxis of MHCC97H cells and HSCs was observed by cell-cell contact method,fibrinogen-thrombin paste technique and in cancer-bearing mice.The proliferation and migration abilities of HSCs were significantly enhanced.Conclusion Hepatoma carcinoma cells can promote the activation,proliferation and migration of HSCs under the cocultured system in vitro and in vivo.