目的研究过表达Tenomodulin(TNMD)对NIH3T3和C3H10T1/2细胞系肌腱相关分子表达的作用。方法体外培养NIH3T3和C3H10T1/2细胞系,利用脂质体法在两种细胞系中分别转染pCAGGS空载体和pCAGGS-TNMD表达载体,G418进行稳定筛选。RT-PCR确认成功转染后,利用倒置相差显微镜观察细胞形态,同时定量PCR检测肌腱相关基因的表达情况。结果两种细胞系转染后细胞形态均无明显改变。基因水平上,NIH3T3细胞过表达TNMD后,collagenⅥ和biglycan显著降低,collagenⅠ没有显著差别。而在C3H10T1/2细胞中,collagenⅠ和biglycan显著升高,collagenⅥ没有显著差别。结论过表达TNMD对NIH3T3和C3H10T1/2细胞系的细胞形态没有影响,但能影响肌腱相关基因的表达情况。

Objective To study the effect of tenomodulin(TNMD) over-expression on the expression of tenocyte related molecules of NIH3T3 and C3H10T1/2 cell lines.Methods The NIH3T3 and C3H10T1/2 cells were cultured in vitro and then transfected with pCAGGS vector and pCAGGS-TNMD expression vector respectively.After confirmation of successful transfection by RT-PCR,the morphology of transfected cells were observed with inverted phase contrast microscope,and the expression of some tenocyte related genes in transfected cells were analyzed with Quantitative PCR.Results There were no obvious changes in the morphology of NIH3T3 and C3H10T1/2 cells after transfection.The gene expression of collagenⅥ and biglycan in NIH3T3 cells decreased dramatically,while that of collagenⅠdid not shown obvious change.However,in C3H10T1/2 cells,the expression of collagenⅠand biglycan increased dramatically respectively,collagenⅥ did not shown obvious difference.Conclusion Tenomodulin(TNMD) over-expression of NIH3T3 and C3H10T1/2 cell lines did not affect the cell morphology,but could affect the expression of some tenocyte related genes.