Sox6基因和Col2a1在大鼠bMSCs体外成软骨分化中表达及其意义  

Expression and significance of Sox6 and Col2a1 during in vitro chondrogenic differentiation of rat bMSCs

作　　者：钱晓伟[1] 谭湘陵[1] 张贤 蔡建平 

机构地区：[1]南通大学生物工程研究所,江苏省226019 [2]无锡中医医院骨伤科

出　　处：《江苏医药》2012年第19期2240-2243,F0002,共5页Jiangsu Medical Journal

基　　金：国家中医药管理局(06-07JP21);南通大学校级课题(092013)

摘　　要：目的探讨大鼠骨髓间充质干细胞(bMSCs)成软骨分化中Sox6基因和Ⅱ型胶原α1(Col2α1)表达水平的变化及其意义。方法 2个月龄SD大鼠分离bMSCs,置含15%胎牛血清的DMEM/F12(1∶1)培养基中培养。传至第3代后,细胞分为诱导组和未诱导组,诱导组在培养基中添加含转化生长因子β1(TGF-β1)的诱导剂,分别在诱导后3、6、12、24h、3、7和14d提取细胞总RNA,逆转录成cDNA,荧光定量PCR法检测Sox6基因和Col2α1基因的表达。同时应用细胞免疫化学法检测诱导14d的Col2α1的表达。结果与未诱导组细胞比较,bMSCs在TGF-β1的诱导下,形态上发生软骨样细胞变化;诱导后6h,Sox6基因的表达上升6.3倍(P<0.01),随后迅速下降,3-14d维持在4.0-4.7倍水平;Col2α1基因的表达量在诱导后3d达1.9倍,7d和14d分别达14.3和35.8倍(P<0.01)。诱导组可见大量Ⅱ型胶原阳性细胞。结论 bMSCs在体外可以分化为软骨样细胞,Sox6基因在bMSCs的成软骨分化的早期和晚期都发挥了一定的作用。Objective To investigate the expression and significance of Sox6 and type Ⅱ collagen al（Col2αl） during in vitro chondrogenic differentiation of rat bone mesenchymal stem cells （bMSCs）. Methods The bMSCs were obtained from femur of 2 months age SD rats and cultured in media of DMEM/F12 （1 ： 1） containing 15%FBS at 37℃, 5% CO2. When bMSCs were cultured in vitro for 3 passages, the cells were seeded to 6-well culture plate and divided into two groups of A （induced by TGF-β1） and B（uninduced as the control）. After induced for 3,6,12,24 h,3,7 and 14 d, total RNA of the cultured cells was extracted and the expressions of Sox6 and Col2αl were detected by RT-PCR. Meanwhile the immunohistoehemical test was used to detect the expression of Co12~1 aftei induced for 14 days. Results Compared to group B, the bMSCs in group A showed the shape ot cartilage cells. The expression of Sox6 was upregulated to 6. 3 times at 6 h（P〈0. 01）, which then declined rapidly and maintained 4. 0 to 4. 7 times 3 days later（P〈0. 01）. The expression of Col2αl was upregulated to 1.9 times at 3 d,14. 3 times at 7 d, and 35.8 times at 14 d（P〈O. 01）. At 14 d, many type 11 collagen positive cells were discovered in group A. Conclusion With the induction ot TGF-β1, bMSCs can differentiate into chondrocytes. Sox6 gene plays a certain role in the early and late stage during chondrogenic differentiation of bMSCs.

关 键 词：骨髓间充质干细胞 成软骨分化 Ⅱ型胶原α1 Sox6基因 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]