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Effects of Targeting Regulation of AKT/mTOR Signaling Pathway on Proliferation， Apoptosis， Autophagy and Osteogenic Differentiation of Osteosarcoma Cell Line MG63
Zhang Yao1, Kang Quan2, Dong Zixing1, Xie Shengnan1, Luo Qing1*
1Pediatric 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; 2Department of Hepatology, Children’s Hospital of Chongqing Medical University, Chongqing 400014, China
Abstract: 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 mRNA expression of AKT and mTOR 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 mTOR 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.
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