靶向调控AKT/mTOR信号通路对骨肉瘤细胞MG63增殖、凋亡、自噬及成骨分化的影响  

Effects of Targeting Regulation of AKT/mTOR Signaling Pathway on Proliferation, Apoptosis, Autophagy and Osteogenic Differentiation of Osteosarcoma Cell Line MG63

作　　者：张遥 康权[2] 董姿杏 谢圣男 罗庆[1] Zhang Yao;Kang Quan;Dong Zixing;Xie Shengnan;Luo Qing(Pediatric Research Institute,Children＇s Hospital of Chongqing Medical University,Ministry of Education Key Laboratory of Child Development and Disorders,China International Science and Technology Cooperation Base of Child Development and Critical Disorder,Lab of Pediatrics Oncology,Chongqing Key Laboratory of Pediatrics,Chongqing 400014,China;Department of Hepatology,Children＇s Hospital of Chongqing Medical University,Chongqing 400014,China)

机构地区：[1]重庆医科大学附属儿童医院儿科研究所儿童发育疾病研究教育部重点实验室儿童发育重大疾病国家国际科技合作基地儿科学重庆市重点实验室,重庆400014 [2]重庆医科大学附属儿童医院肝胆外科,重庆400014

出　　处：《中国细胞生物学学报》2018年第9期1486-1493,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81172545);重庆市卫生和计划生育委员会医学科研项目(批准号:2016MSXM093)资助的课题~~

摘　　要：该文探究靶向调控AKT/mTOR信号通路后,对人骨肉瘤细胞株(MG63)增殖、凋亡、自噬及成骨分化的影响并探讨其机制。RT-PCR检测在不同恶性程度骨肉瘤细胞中AKT、mTOR基因表达的情况;选择靶向mTOR信号通路的抑制剂雷帕霉素和激活剂3-苄基-5-(2-硝基苯氧甲基)-γ-丁内酯(3-BDO),分别用CCK-8检测细胞增殖;DAPI染色、Annexin V-FITC/PI双染法检测凋亡;碱性磷酸酶(ALP)染色检测早期成骨能力;茜素红染色检测中晚期成骨能力;Western blot技术检测自噬相关蛋白及分化抑制因子(Id1)表达。结果显示,AKT/mTOR表达情况与骨肉瘤恶性程度有关;通过靶向抑制AKT/mTOR信号通路后,可抑制骨肉瘤细胞MG63增殖,促进凋亡,上调自噬水平,抑制其早、晚期成骨分化;靶向激活AKT/mTOR信号通路后,对骨肉瘤细胞MG63增殖、凋亡无明显影响,下调自噬水平,但可促进其早、晚期成骨分化。该研究表明,靶向调控AKT/mTOR信号通路与分化抑制因子(Id1)表达有关,可进一步阐明骨肉瘤发病机制,为诱导分化治疗提供理论依据。The aim of this paper was to investigate the effects of targeted regulation of AKT/mTOR signaling pathway on the proliferation, apoptosis, autophagy and osteogenic differentiation of human osteosarcoma cell line（MG63）, and to explore its mechanism. RT-PCR was used to detect m RNA expression of AKT and m TOR in osteosarcoma cells with different degrees of malignancy. Rapamycin was used as a targeted inhibitor of the mtor signaling pathway, and 3-BDO was used as a targeted activator of the mtor signaling pathway. Cell proliferation activity was detected by CCK-8 assay, apoptosis was detected by DAPI staining and Annexin V-FITC/PI double staining; alkaline phosphatase（ALP） staining was used to detect cell early osteogenic differentiation ability. Alizarin red staining was used to detect cell middle-late osteogenic differentiation ability. Autophagy and Id1 was detected by Western blot. The results showed that expression of AKT/mTOR was related to the degree of malignancy of osteosarcoma. After targeted inhibition of AKT/mTOR signaling pathway, inhibitor of m TOR signaling pathway could inhibit the proliferation of osteosarcoma cell MG63 and promote apoptosis, upregulate autophagy, but inhibite early and late osteogenic differentiation. After targeted activation of AKT/mTOR signaling pathway, there was no significant effect on the proliferation and apoptosis of osteosarcoma cell MG63, down-regulated autophagy, but promoted its early and late osteogenic differentiation. This study indicated that targeted regulation of AKT/mTOR signaling pathway was related to the expression of differentiation suppressor factor（Id1）, which could further elucidate the pathogenesis of osteosarcoma and provide theoretical basis for inducing differentiation therapy.

关 键 词：AKT/MTOR 骨肉瘤 增殖 凋亡 自噬 成骨分化 

分 类 号：R738.1[医药卫生—肿瘤]