骨髓和肝癌来源间充质干细胞对肝癌细胞HepG2增殖、侵袭能力的影响

The effects of bone marrow mesenchymal stem cells and liver cancer derived mesenchymal stem cells on the proliferation and invasion of HepG2 cells

目的 观察骨髓间充质干细胞（BMSCs）和肝癌来源间充质干细胞（LCMSCs）对肝癌细胞HepG2增殖、侵袭能力的影响.方法 贴壁筛选法分离培养LCMSCs,成脂与成骨诱导分化鉴定其分化能力,流式细胞仪检测其表面标志物.分别收集BMSCs和LCMSCs培养上清,与HepG2细胞共培养,以完全培养基为对照组,细胞计数试剂盒（CCK-8）法和Transwell侵袭实验检测各组细胞增殖与侵袭能力.结果 分离的LCMSCs形态均一,具有良好的成脂与成骨分化能力,阳性表达CD29和CD105,阴性表达CD34和CD45.CCK-8法证实BMSCs和LCMSCs培养上清均能促进HepG2细胞增殖,与BMSCs培养上清比较,LCMSCs培养上清对HepG2细胞增殖的促进作用更明显（P＜0.05）.Transwell侵袭实验表明,对照组、BMSCs培养上清组及LCMSCs培养上清组穿膜细胞数分别为13、24、35个,BMSCs培养上清和LCMSCs培养上清处理后,HepG2细胞的侵袭能力明显增强;与BMSCs CM比较,LCMSCs CM对HepG2侵袭能力的增强作用更为明显（P＜0.05）.结论 BMSCs和LCMSCs均可增强肝癌细胞HepG2的增殖和侵袭能力,但LCMSCs对HepG2细胞的作用更强.

Objective To investigate the effects of bone marrow mesenchymal stem cells （BMSCs） and liver cancer derived mesenchymal stem cells （LCMSCs） on the proliferation and invasion of HepG2 cells.Methods LCMSCs were isolated and cultured by adherent screening,and identified by adipogenic and osteogenic induction and flow cytometry.HepG2 cells were treated with conditioning medium collected from BMSCs or LCMSCs （BMSCs CM or LCMSCs CM）,and complete medium as control,the changes of proliferation and invasion of HepG2 cells were tested by a cell counting kit-8 （CCK-8） assay and transwell chambers respectively.Results Cultured LCMSCs presented in a uniform spindle morphology,had a good ability of adipogenic and osteogenic differentiation,and were positive for CD29 and CD105,negative for CD34 and CD45.CCK-8 results showed that BMSCs CM or LCMSCs CM significantly promoted proliferation in vitro,and the proliferative effect in LCMSCs CM group was more obvious than that in BMSCs CM group （P 〈 0.05）.The Transwell chambers results revealed that cell invasion was significantly enhanced after cells treated with BMSCs CM or LCMSCs CM,the number of penetrating cells in control group,BMSCs CM group and LCMSCs CM group was 13,24 and 35 respectively,and the invasive effect in LCMSCs CM group was more obvious than in BMSCs CM group （P 〈 0.05）.Conclusion BMSCs and LCMSCs can enhance the proliferation and invasion of HepG2 cells,and the proliferative and invasive effects of LCMSCs seem stronger.