人骨髓间充质干细胞对肝癌细胞的生物学效应  

Biological effect of human bone marrow mesenchymal stem cells on hepatocellular carcinoma cells

作　　者：陈琪枫[1] 方晓明[1] 姚宁[1] 方旭东[1] 龚谋春[1] 

机构地区：[1]解放军第117医院普外科,浙江省杭州市310013

出　　处：《中国组织工程研究》2015年第10期1511-1515,共5页Chinese Journal of Tissue Engineering Research

摘　　要：背景:肝癌的转移特性是影响预后的关键因素,观察人骨髓间充质干细胞对肝癌转移行为的影响,对于提高肝癌患者的生存期具有重要意义。目的:探讨人骨髓间充质干细胞对不同转移潜能肝癌细胞生物学特性的影响。方法:Transwell下室加入人骨髓间充质干细胞,上室加入高、低转移潜能肝癌细胞悬液,共培养36 h酶标仪检测吸光度值及细胞计数法观察肝癌细胞侵袭能力的变化。CCK-8法检测人骨髓间充质干细胞对高低转移潜能肝癌细胞增殖能力的影响。PCR检测人骨髓间充质干细胞与高低转移潜能肝癌细胞共培养前后转移相关因子骨桥蛋白、唾液酸蛋白、整合(α-V)以及增殖相关基因转化生长因子β1和程序化凋亡因子的表达。结果与结论:1共培养组的肝癌细胞迁移数量显著少于单独培养组,经酶标仪半定量检测肝癌细胞侵袭性显示,共培养组吸光度值明显低于单独培养组,差异有显著性意义(P<0.05)。2高转移潜能肝癌细胞与人骨髓间充质干细胞共培养组转移相关因子骨桥蛋白和骨唾液酸蛋白表达显著下降(P<0.05),但整合(α-V)基因表达未出现明显改变(P>0.05)。低转移潜能肝癌细胞与人骨髓间充质干细胞共培养组转移相关因子骨桥蛋白、骨唾液酸蛋白以及整合(α-V)基因表达显著下降(P<0.05)。3经酶标仪半定量检测肝癌细胞增殖能力显示,共培养组吸光度值明显高于单独培养组,差异有显著性意义(P<0.05)。4高转移潜能肝癌细胞与人骨髓间充质干细胞共培养组的转化生长因子β1基因表达显著上调(P<0.05);但程序化凋亡因子基因表达无明显改变(P>0.05)。低转移潜能肝癌细胞与人骨髓间充质干细胞共培养组的转化生长因子β1、程序化凋亡因子基因表达均显著上调(P<0.05)。以上结果提示人骨髓间充质干细胞与肝癌细胞共培养后,肝癌细胞侵袭能力明显下调,增殖能力明显提高。BACKGROUND： The metastatic potential of hepatocellular carcinoma cells is key factor influencing patient＇s prognosis. To observe the effect of human bone marrow mesenchymal stem cells on metastasis of hepatocellular carcinoma is of great significance for improving the lifetime of hepatocellular carcinoma patients. OBJECTIVE： To explore the biological effect of human bone marrow mesenchymal stem cells on hepatocellular carcinoma cells with different metastatic potentials. METHODS： Human bone marrow mesenchymal stem cells and hepatocellular carcinoma cell suspension with high and low metastatic potentials were respectively injected into the Transwell chamber, and after 36 hours of co-cuifure, ELISA method was used to detect the absorbance value as welt as cell counting method was used to observe the changes in the invasion ability of hepatocellular carcinoma cells. The effects of human bone marrow mesenchymal stem cells on the proliferation of hepatocellular carcinoma cell suspension with high and low metastatic potentials were determined using cellcounting kit-8. PCR method was adopted to measure the expression of osteopontin, bone specific sialoproteins, integration （alpha V）, transforming growth factor beta 1 and programmed cell death protein 5. RESULTS AND CONCLUSION： （1） The number of migrated hepatocellular carcinoma cells was significantly lower in the co-culture group than the single culture group, and based on the semi-quantitative detection of invasion ability, the absorbance value of the co-culture group was significantly lower than that in the single culture group （P 〈 0.05）. （2） The expression of osteopontin and bone specific sialoproteins was significantly decreased in the co-culture group with high metastatic potential （P 〈 0.05）, but there was no change in the expression of integration （alpha V） （P 〉 0.05）. In the co-culture group with low metastatic potential, the expression of osteopontin, bone specific sialoproteins, and integration （alpha V） were d

关 键 词：骨髓 间质干细胞 癌 肝细胞 肿瘤侵润 干细胞 骨髓干细胞 肝癌细胞 人骨髓间充质干细胞 生物学行为 侵袭能力 细胞增殖 

分 类 号：R394.2[医药卫生—医学遗传学]