EMILIN3基因促进人骨髓间充质干细胞增殖及向软骨细胞分化

EMILIN3 stimulates human mesenchymal stem cells proliferation and differentiation into cartilage cells in vitro

目的探讨EMILIN3基因对人骨髓间充质干细胞(human marrow mesenchymal stem cells,hMSCs)增殖及向软骨细胞分化的影响。方法构建EMILIN3腺病毒表达载体,细胞分成hMSCs、hMSCs+对照腺病毒、hMSCs+EMILIN3腺病毒3组,MTT法检测细胞的增殖情况,Real-time PCR法检测上述细胞向软骨细胞诱导后CollagenⅡ、Aggrecan、SOX9mRNA的表达。结果hMSCs+EMILIN3腺病毒组细胞增殖能力明显快于hMSCs+对照腺病毒组和hMSCs组(P<0.05),hMSCs+对照腺病毒组与hMSCs组增殖能力相比有所降低,但差异无统计学意义(P>0.05)。hMSCs+EMILIN3腺病毒组细胞中CollagenⅡ、Aggrecan、SOX9mRNA的相对表达量与hMSCs+对照腺病毒组、hMSCs组相比明显增加(P<0.05,P<0.01)。hMSCs+对照病毒组与hMSCs组相比有所增加,但没有明显的趋势(P>0.05)。结论EMILIN3促进hMSCs增殖,并且可以增加hMSCs向软骨细胞诱导分化。

Objective To determine the effect of EMILIN3 gene on the proliferation of human marrow mesenchymal stem cells （hMSCs） and the differentiation into cartilage cells. Methods The adenovirus plasmid of EMILIN3 was constructed. The cells were transfected with the plasmid or the blank vector, and the cells without transfection served as control. The proliferation of these cells was detected by MTT assay. The expression of CollagenⅡ, Aggrecan and SOX9 at mRNA level was also detected with real-time PCR. Results The proliferation was significantly increased in the hMSCs infected by the adenovirus plasmid of EMILIN3 than the cells with blank vector infection and control cells （P〈0.05）. But there was no difference in the proliferation between the 2 later groups of cells （P〉0.05）. The expression of CollagenⅡ, Aggrecan and SOX9 was increased by 2.5, 2.7 and 2.1 times respectively in the hMSCs infected by the adenovirus with EMILIN3 than that in the hMSCs infected by the control adenovirus （P〈0.05）. Compared with the hMSCs, there was no difference in the expression of above genes in the hMSCs infected by the blank vector. Conclusion EMILIN3 transfection enhances hMSCs proliferation and stimulate the cells to differentiate into cartilage cells.