中国组织工程研究Issue(50)：8056-8061,6.DOI:10.3969/j.issn.2095-4344.2015.50.004
共沉默miR-221-3p/222-3p表达抑制骨髓间充质干细胞增殖及促成软骨分化
Down-regulation of miR-221-3p/222-3p inhibits cell proliferation and promotes chondrogenic differentiation of human bone marrow mesenchymal stem cells
摘要
Abstract
BACKGROUND:The use of mesenchymal stem cels in the field of tissue engineering for osteoarticular injury repair is a very promising tool since these cels are readily expandable and able to differentiate into chondrocytes. Abundant evidence suggests that microRNAs play critical roles in chondrogenic differentiation of mesenchymal stem cels. <br> OBJECTIVE:To observe the chondrogenic effect of human bone marrow mesenchymal stem cels transfected with lentiviral vectors bearing miR-221-3p/222-3p inhibition, thereby provding new strategies for cartilage injury. <br> METHODS: miRNA microarray technology was applied to detect microRNAs expression profiles at three different stages of chondrogenic differentiation induction after transforming growth factor-β3 treatment and verified by real-time fluorescence quantitative PCR (RT-qPCR). Human bone marrow mesenchymal stem cels were infected with lentivirus bearing miR-221-3p/222-3p inhibition. After co-suppressing the expression of miR-221/222-3p, cel counting kit-8 was used to determine the cel proliferation, the differentiation of bone marrow mesenchymal stem cels towards chondrocytes was verified by type II colagen protein expression through immunohistochemistry and glycosaminoglycan accumulation was also elevated by sarranine O staining. RT-PCR was used to detect type II colagen and aggrecan mRNA expression at 21 days of chondrogenic induction. <br> RESULTS AND CONCLUSION: The expression of miR-221-3p/222-3p was inhibited after Lv-miR221-3p/222-3p inhibition co-transfected into bone marrow mesenchymal stem cels. microRNA microarray and RT-qPCR results showed that the expression of miR-221-3p/222-3p was declined significantly at the anaphase of chondrogenic differentiation. The expression levels of chondrogenic markers, Aggrecan and type II colagen were significantly increased in the miR-221-3p/222-3p inhibition group and cel proliferation was also inhibited significantly compared with non-transduced cels or transduced with the empty lentiviral vector group. miR-221-3p/222-3p knockdown in bone marrow mesenchymal stem cels could inhibit proliferation but promote chondrogenic differentiation of bone marrow mesenchymal stem cels.关键词
干细胞/骨髓干细胞/成软骨分化/miR-221-3p/miR-222-3p/慢病毒表达载体/骨髓间充质干细胞/沉默miRNA/国家自然科学基金Key words
Bone Marrow/Mesenchymal Stem Cels/Cel Differentiation/MicroRNAs/Tissue Engineering分类
医药卫生引用本文复制引用
闫继红,周德山,杨姝,孙海梅,曹丹丹,张秀英,季凤清,郭多,吴波,孙婷怡..共沉默miR-221-3p/222-3p表达抑制骨髓间充质干细胞增殖及促成软骨分化[J].中国组织工程研究,2015,(50):8056-8061,6.基金项目
国家自然科学基金面上项目(81271728)；北京市自然科学基金面上项目(7152023)Funding：the National Natural Science Foundation of China, No.81271728 （81271728）
the Natural Science Foundation of Beijing, No.7152023 （）
