Sox9基因转染诱导L6大鼠成肌细胞向软骨细胞表型分化的研究

The research on Sox9 gene transfection induced chondrogenic differentiation of L6 rat myoblasts

利用脂质体转染法,将真核表达质粒pcDNA3.1-Sox9导入L6细胞中.通过细胞增殖检测、细胞形态学观察、标志基因表达分析和甲苯胺蓝染色等方法分析Sox9基因转染对L6细胞增殖和向成软骨方向分化的诱导作用.结果表明,Sox9基因转染对L6细胞的增殖能力没有影响,但能明显抑制L6细胞相互融合形成肌管.和对照相比,Sox9基因转染后,细胞成肌分化标志基因Myf5的表达受到明显抑制,而成软骨分化标志基因Ⅱ型胶原(typeⅡcollagen,Col2a1)和蛋白聚糖(Aggrecan,Agg)的表达显著提高.转染后14 d,Sox9基因转染的细胞可见甲苯胺蓝染色阳性的软骨细胞样结节,而对照组则鲜见类似结构.上述结果表明,Sox9基因转染可以抑制L6细胞向成熟肌细胞分化,并促进其向软骨表型分化.

L6 myoblasts were transfected with the eucaryotic expression plasmid pcDNA3. 1-5ox9 using Lipofectamine 2000. The proliferation and chondrogenie differentiation of L6 cells was determined by cell proliferation assay, cell morphology examination, evaluation of marker gene expression and toluidine blue staining. It was shown that Sox9 gene transfer did not influence L6 cell proliferation; however, it could significantly inhibit myotube formation. Compared with the control group, the expression of myogenic maker My f5 was remarkably suppressed in the Sox9 group, whereas the expression of chondrogenic markers type 11 collagen （Col2al） and Aggrecan （Agg） was significantly enhanced. 14 days after transfection ,cartilage-like nodules with positive toluidine blue stain were readily observed in the Sox9 group but rarely found in the control group. The above results indicated that Sox9 gene transfection could disrupt the myogenesis of L6 cells and induce them to differentiate towards chondrogenic phenotype.