骨髓间充质干细胞对HepG2肝癌细胞增殖的影响
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江苏省科技支撑计划资助项目(BE2011842)


The effects of bone marrow-derived mesenchymal stem cells on proliferation of human hepatocellular carcinoma cell line HepG2 in vitro
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美国Cure Cancer Worldwide corporation资助项目

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    摘要:

    目的:观察大鼠骨髓间充质干细胞对HepG2肝癌细胞增殖的影响,并初步探讨其机制-方法:全骨髓贴壁法分离培养大鼠骨髓间充质干细胞,流式细胞术鉴定其分子表面标志,体外扩增培养并收集培养上清-MTT法-平板克隆实验及软琼脂克隆实验检测大鼠间充质干细胞对人肝癌细胞株HepG2增殖的影响,流式细胞术检测肝癌细胞周期的改变,Western blot法检测细胞周期相关蛋白Cyclin D1表达情况-结果:骨髓间充质干细胞CD105和CD90表达阳性,CD34和CD45表达阴性-在骨髓间充质干细胞培养上清作用下,MTT法显示实验组肝癌细胞光密度值增加(P < 0.05),平板克隆法显示实验组肝癌细胞克隆形成率比对照组明显升高(P < 0.05),软琼脂克隆法显示实验组肝癌细胞空间克隆形成率比对照组明显升高(P < 0.05),流式细胞术显示实验组细胞周期G1期比例降低,S期-G2期比例增加-Western blot显示实验组细胞Cyclin D1表达量增加(P < 0.05)-结论:骨髓间充质干细胞可促进HepG2肝癌细胞的增殖及空间克隆形成能力,其促进肝癌细胞体外增殖可能与Cyclin D1表达上调有关-

    Abstract:

    Objective: To investigate the effects of rat bone marrow-derived mesenchymal stem cells (MSCs) on the proliferation and cell cycle distribution of human hepatocellular carcinoma cell line HepG2 in vitro. Methods: Rat bone marrow-derived mesenchymal stem cells were isolated and identified by FACS and were expanded ex vivo and the supernatant was collected. Cell growth was measured by MTT assay,colony formation and soft agar colony formation assay. Cell cycle progression was analyzed by flow cytometry. Western blot assays were used to detect the expression of Cyclin D1. Results: Mesenchymal stem cells promoted the proliferation and clonogenicity of cell line HepG2 indirectly by MSC-CM(P < 0.05). Cell number in G0/G1 phase was decreased and that in S and G2/M phase was increased(P < 0.05). MSCs promote the expression of protein Cyclin D1 in HepG2(P < 0.05). Conclusion:Mesenchymal stem cells can promote the proliferation of hepatocellular carcinoma cell line by upregulating Cyclin D1.

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唐 甜,李 红,薛 冰,唐小军,陈汐敏,仇镇宁,朱 进,Tom C Tsang,冯振卿.骨髓间充质干细胞对HepG2肝癌细胞增殖的影响[J].南京医科大学学报(自然科学版),2012,(5):604-608

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  • 收稿日期:2011-12-29
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