Zheng Chang-Jun, Yang Li-Li, Liu Jun, Zhong Lei
a Department of Orthopaedics , The 2nd Hospital of Jilin University , Changchun , PR China.
b Department of Spine Surgery , The 2nd Hospital of Jilin University , Changchun , PR China.
J Recept Signal Transduct Res. 2018 Apr;38(2):133-140. doi: 10.1080/10799893.2018.1436561. Epub 2018 Feb 15.
The research of G protein-coupled receptors (GPCRs) is a promising strategy for drug discovery. In cancer therapy, there is a need to discover novel agents that can inhibit proliferation and induce apoptosis in cancer cells. JTC-801 is a novel GPCR antagonist with the function of reversing pain and anxiety symptoms. This study aims to investigate the antitumor effects of JTC-801 on human osteosarcoma cells (U2OS) and elucidate the underlying mechanism.
The Cell Counting Kit-8 assay was used to detect the viability of U2OS cells treated with JTC-801 in vitro. The cell apoptosis was evaluated using a flow cytometry assay with Annexin V-FITC/PI double staining. The inhibitory effect of JTC-801 on invasion and migration of U2OS cells were determined by the Transwell assays. Western blot assay was performed to measure the levels of proteins related to cell apoptosis and its mechanism.
The JTC-801 significantly decreased the viability of U2OS cells (p < .05) as a result of its anti-proliferative effect through induction of apoptosis associated with activation of BAX, Caspase-3 and down-regulating BCL-2 expression. The invasive and migratory cells were obviously reduced after JTC-801 treatment (p < .05). Further, the phosphorylated AKT, mTOR and active p70 S6 protein kinase in the PI3K/AKT signaling pathway were obviously lessened in the JTC-801 treated U2OS group (p < .05).
JTC-801 may exert osteosarcoma cell growth inhibition by promoting cell apoptosis, through PI3K/AKT signaling pathway participation.
G蛋白偶联受体（GPCRs）的研究是药物研发的一个有前景的策略。在癌症治疗中，需要发现能够抑制癌细胞增殖并诱导其凋亡的新型药物。JTC-801是一种新型GPCR拮抗剂，具有逆转疼痛和焦虑症状的功能。本研究旨在探讨JTC-801对人骨肉瘤细胞（U2OS）的抗肿瘤作用，并阐明其潜在机制。
采用细胞计数试剂盒-8法检测体外JTC-801处理的U2OS细胞的活力。使用Annexin V-FITC/PI双染流式细胞术检测细胞凋亡。通过Transwell实验测定JTC-801对U2OS细胞侵袭和迁移的抑制作用。进行蛋白质印迹分析以测量与细胞凋亡及其机制相关的蛋白质水平。
JTC-801通过诱导与BAX、Caspase-3激活相关的凋亡并下调BCL-2表达，发挥抗增殖作用，显著降低了U2OS细胞的活力（p＜0.05）。JTC-801处理后，侵袭和迁移细胞明显减少（p＜0.05）。此外，在JTC-801处理的U2OS组中，PI3K/AKT信号通路中磷酸化的AKT、mTOR和活性p70 S6蛋白激酶明显减少（p＜0.05）。
JTC-801可能通过PI3K/AKT信号通路参与促进细胞凋亡，从而发挥对骨肉瘤细胞生长的抑制作用。
