摘要
目的:观察细胞因子诱导成骨的共同信号分子核心结合因子α1基因转染NIH3T3成纤维细胞,其成骨表型转变的可能性。方法:实验于2003-10/2004-10在解放军第三军医大学新桥医院骨科中心实验室完成。采用脂质体转染技术行核心结合因子α1基因转染NIH3T3成纤维细胞,成骨标志基因I型胶原、骨钙素、骨唾液蛋白、碱性磷酸酶、核心结合因子α1mRNA的表达采用反转录-聚合酶链反应检测,骨钙素的表达采用免疫细胞化学染色检测,Cbfa蛋白的表达采用Westernblot检测。结果:①NIH3T3成纤维细胞经核心结合因子α1基因转染后出现骨钙素、骨唾液蛋白、碱性磷酸酶的表达,I型胶原表达增强,说明在转录水平有成骨标志基因的表达。②免疫细胞化学染色检测到了骨钙素的表达,显示NIH3T3细胞已发展为成骨细胞。③Westernblot检测到了核心结合因子α1蛋白的表达,证实了成纤维细胞发生了表型的改变。结论:核心结合因子α1基因转移NIH3T3成纤维细胞改变了成纤维细胞向成骨表型转化,增加了成骨细胞数量,使成骨能力增强成为可能。
AIM： To observe the fibroblast NIH3T3 transfected by the same signal molecule core -binding factor α1 （Cbfal） gene of the osteogenesis induced by cytokine and the possibility of changes of its osteogenesis phenotype. METHODS：The experiment was done in the Central Laboratory of Department of Orthopaedics of Xinqiao Hospital of Third Military Medical University of Chinese PLA from October 2003 to October 2004. The gene trasfection of Cbfal to fibroblast NIH3T3 was performed with liposomes trasfection technique. The expression of osteogenesis representative gene： I Collagen, Osteocalcin, bone sialoprotein （BSP）, alkaline phosphatase （AKP）, and Cbfal mRNA were detected with reverse transcription-polymerase chain reaction （RTPCR）. The expression of osteocalcin was detected with immunocytochemical stain. The expression of Cbfa protein was detected with Westernblot. RESULTS： ① The expression of osteocalcin, bone sialoprotein and alkaline phosphatase appeared after the trasfection of Cbfal gene to fibroblast NIH3T3. The reinforcement of I type collagen expression indicated that the expression of osteogenesis representative gene existed in the transcription level. ② The expression of osteocalcin was detected by immunocytochemical stain, which showed that the NIH3T3 cells had developed the osteogenesis cells. ③ The expression of Cbfal protein was detected by Westernblot, which confirmed that the phenotype changes had occurred on fibroblast. CONCLUSION： Gene transfer of Cbfal to fibroblast NIH3T3 can alter the conversion of the phenotype of osteogenesis, and then the quantity of osteoblast increased and made the increase of osteogenesis ability possible.
出处
《中国临床康复》
CSCD
北大核心
2005年第26期118-120,共3页
Chinese Journal of Clinical Rehabilitation
