骨髓间充质干细胞对高低转移潜能肝癌细胞影响的实验研究

目的观察人骨髓间充质干细胞(hMSC)对高低转移潜能肝癌细胞(MHCC97-H和MHCC97-L)侵袭能力和增殖能力的影响。方法 Transwell法检测hMSC对高低转移潜能肝癌细胞侵袭能力的影响,下室加入培养基和hMSC,上室加入高低转移潜能肝癌细胞悬液,共培养36 h,酶标仪吸光度(optical density,OD)值检测及细胞计数法观察侵袭能力的变化。CCK-8法检测hMSC对高低转移潜能肝癌细胞增殖能力的影响。PCR检测共培养前后转移相关因子骨桥蛋白(OPN)、唾液酸蛋白(BSP)、α-V基因、增殖相关基因TGFβ1和PDCD4的表达差异。结果 (1)侵袭能力检测结果:与对照组比较,镜下观察实验组高低转移潜能肝癌细胞迁移数量明显减少,OD值两组比较差异有统计学意义(P0.05)。MHCC97-H与hMSC共培养后OPN、BSP表达明显下降(P0.05),而α-V表达无明显变化(P0.05)。MHCC97-L与hMSC共培养后OPN、BSP、α-V表达均显著下降(P0.05)。(2)增殖能力检测结果:与对照组比较,实验组高低转移潜能肝癌细胞OD值明显增高(P0.05),实验组TGFβ1基因表达上调(P0.05),而PDCD4基因表达在两组间差异无统计学意义(P0.05),在MHCC97-L实验组PDCD4基因表达上调(P0.05)。结论 hMSC使高低转移潜能肝癌细胞的侵袭能力下调,增殖能力增强。

Experiment study of hMSC affecting high and low metastatic potential hepatocelluar carcinoma cells

Objective To investigate the invasion and proliferation ability changes of HCC cells with different metastatic potential by hMSC transplantation.Methods Transwell assay was used to detect the effects of hMSC on invasion ability of human HCC cells.The lower chamber was added medium and hMSC,and upper chamber was added human HCC cells,hMSC and HCC cells were co-cultured 36 hours in incubator.Invasive ability changes of HCC cells was analyzed by cells count results and optical density （OD）value of sample cells by microplate reader.The effect of hMSC on HCC cells proliferation ability was detected by CCK-8 assay.The expression of the metastatic and proliferative related factors genes of HCC cells before and after co-culture with hMSC by PCR were detected.Results （1） Invasion ability test results：compared with control group,morphologic study showed HCC cells migration quantity was decreased significantly experimental group,and OD value was significant difference between the two groups （P ＜ 0.05）.OPN,BSP gene expression of MHCC97-H cells showed a significant decline in the co-culture with hMSC group compared with the MHCC97-H control one （P ＜0.05）,and α-Ⅴwas no significant difference （P ＞0.05）.OPN,BSP,α-Ⅴ gene expression of MHCC97-L cells showed a significant decline in the co-culture with hMSC group compared with the MHCC97-L control one （P ＜0.05）.（2） Proliferation ability test results：compared with control group,OD value was increased significantly in two experimental groups （P ＜0.05）,and TGFβ1 gene expression was up-regulation in two experimental groups （P ＜ 0.05）.PDCD4 gene expression was no significant difference between MHCC97-H cell co-culture with hMSC group and MHCC97-H cell control group （P ＞ 0.05）.PDCD4 gene expression was significant difference between MHCC97-L cell co-culture with hMSC group and MHCC97-L cell control group （P ＜ 0.05）.Conclusion The experimental results showed the hMSC made the invasion ability of different metastatic po