初级纤毛在多发性骨软骨瘤形成中的作用及其机制  

The studies of roles and mechanism of primary cilium in the progressing of multiple osteochondromas

作　　者：宋登新[1] 曹云[1] 何小文[1] 丁徐[1] 张树威[2] 

机构地区：[1]上海市第一人民医院宝山分院骨科,200940 [2]武汉大学中南医院骨科,430071

出　　处：《中华实验外科杂志》2018年第1期22-25,共4页Chinese Journal of Experimental Surgery

基　　金：上海市卫生计生委员会课题（20124337）

摘　　要：目的比较多发性骨软骨瘤软骨细胞（MOCC）与正常骨骺生长板软骨细胞（NEGPC）初级纤毛的差异，探讨软骨细胞表面的初级纤毛在多发性骨软骨瘤（MO）中的作用。方法培养MOCC和NEGPC，激光共聚焦显微镜观察MOCC和NEGPC表面初级纤毛的形态和发生率；甲苯胺蓝染色及磺酰罗丹明B比色（SRB）法观察MOCC和NEGPC软骨细胞形态及增殖特点；化学染色法比较MOCC和NEGPC两组软骨细胞功能成熟程度[阿尔新蓝和碱性磷酸酶（ALP）染色]的差异。结果体外培养MOCC和NEGPC细胞呈多角形或梭形，甲苯胺蓝染色细胞中出现紫色异染颗粒。免疫荧光染色MOCC细胞表面初级纤毛发生率[（25.265±1.766）%]低于NEGPC细胞[（53.943±3.631）%]，差异有统计学意义（F＝232.692，P＝0.000）。随着细胞生长时间的延长，两组软骨细胞的吸光度（A）值均处于增加状态，但MOCC细胞始终低于NEGPC细胞。组织化学染色MOCC细胞阿尔新蓝染色较浅，而NEGPC细胞着色较深；MOCC细胞ALP染色形成面积[（7.387±2.045） mm2]低于NEGPC[（14.768±2.072） mm2]，差异有统计学意义（F＝16.835，P＝0.020）。结论软骨细胞表面的初级纤毛与MO地形成密切相关，MO软骨细胞表面初级纤毛减少，可能导致骨骺生长板内软骨细胞的增殖、成熟、分泌功能下降。Objective To explore the pathological role of primary cilium in multiple osteochondromas by comparing the difference of primary cilium in the chondrocytes between multiple osteochondromas cartilage cap （MOCC）and normal epiphyseal growth plate cartilage （NEGPC）.MethodsChondrocytes lines from MOCC and NEGPC were collected and cultured in vitro. The morphology and incidence of primary cilla in the MOCC and NEGPC were studied by laser confocal scanning microscope, Toluidine blue staining and Sulforhodamine B （SRB） was applied to display different cell morphology and growth rates between MOCC and NEGPC, and alcian blue and alkaline phosphatase （ALP） in MOCC and NEGPC were measured by histochemical staining.ResultsMOCC and NEGPC cultured in vitro were polygonal or fusiform and were stained with purple particles by toluidine blue staining. The incidence of primary cilla in MOCC [（25.265±1.766）%] was lower than in NEGPC [（53.943±3.631）%], the difference was significant （F=232.692, P=0.000）. A values in MOCC were significantly lower than in NEGPC, while both of them were tended to increase with time. Alcian blue staining in MOCC were decreased compared to NEGPC. The area of ALP staining （7.387±2.045） mm2 in MOCC were decreased obviously compared with NEGPC （14.768±2.072） mm2, the difference was significant （F=16.835, P=0.020）.ConclusionThe primary cilium in chondrocyte was closely related to MO. When the expression of primary cilium in chondrocytes was suppressed, the proliferation, maturation and synthetic function of chondrocytes were delayed in epiphyseal growth plate cartilage.

关 键 词：多发性骨软骨瘤 软骨细胞 初级纤毛 细胞培养 

分 类 号：R738[医药卫生—肿瘤]