SOX6和SOX9基因转染对人原发性骨关节炎关节软骨间充质祖细胞增殖和成软骨分化的调控作用

摘要

Objective To observe the growth and proliferation capabilities of MPCs in primary OA articular cartilage and their differen-tiation properties into chondrocytes by applying related genes SOX6 and SOX9, so as to provide theoretical evidence in preventing and curing primary OA. Methods SOX6 and SOX9 genes were respectively ligated into adenovirus shuttle plasmids pAdTrack-CMV-SOX6 and pAdTrack-CMV-SOX9, then the recombinant plasmids were used to infect MPCs derived from primary OA articular cartilage. TB and the ex-pressions of collagen type Ⅱ protein and mRNA in differentiated MPCs were compared between the infected group and the uninfected group. Results Either SOX6 gene or SOX9 gene could stably infect MPCs from primary OA cartilage. TB and collagen typeⅡwere strongly posi-tive in the SOX6-infected or SOX9-infected MPCs, while they were weekly positive in the uninfected MPCs. Collagen typeⅡmRNA expres-sion in SOX6-infected MPCs derived from primary OA cartilage was 3. 8 times of that in uninfected cells (P<0. 01), and that in SOX9-in-fected MPCs was 5. 15 times of that in the uninfected cells (P<0. 01). Conclusion The stable transfection of SOX6 and SOX9 genes into MPCs derived from primary OA cartilage could significantly promote chondrogenic differentiation of MPCs. There must be feasible methods of gene technology to promote cell proliferation and differentiation of MPCs for repairing articular cartilage injury.%目的：观察SOX6和SOX9基因转染对原发性OA关节软骨MPCs的促增殖、分化作用,为通过调控关节软骨MPCs以防治原发性OA提供理论依据。方法分别以pAdTrack-CMV-SOX6、SOX9腺病毒穿梭质粒构建SOX6、SOX9基因,并感染原发性OA关节软骨MPCs,比较基因感染组和未感染组成软骨诱导分化后 TB、Ⅱ型胶原以及Ⅱ型胶原 mRNA 表达的变化。结果SOX6和SOX9能够分别稳定感染OA关节软骨MPCs；经二者分别感染的关节软骨MPCs成软骨诱导分化后,其TB染色、Ⅱ型胶原染色呈强阳性表达,未基因感染细胞为弱阳性着色；SOX6基因感染原发性OA关节软骨MPCs的Ⅱ型胶原mRNA表达量为未基因感染细胞的3.8倍(P<0.01),SOX9基因为未感染细胞的5.15倍(P<0.01)。结论构建的SOX6、SOX9基因序列与基因库报道序列完全一致；SOX6和SOX9能稳定感染原发性OA关节软骨MPCs,并显著促进感染细胞成软骨分化；提示通过适宜浓度的bF-GF、TGF-β1对原发性OA关节软骨MPCs的作用及SOX6和SOX9基因感染,可能具有促进原发性OA关节软骨损伤修复的作用。