Lin Hang, Hao Yi, Wan Xiaoqing, He Jun, Tong Yongjun
Department of Orthopedics, Zhejiang Hospital, Hangzhou City, China.
J Recept Signal Transduct Res. 2020 Feb;40(1):49-57. doi: 10.1080/10799893.2020.1713807. Epub 2020 Jan 17.
Osteosarcoma is a highly malignant primary tumor. Baicalein has broad-spectrum anti-tumor effects. This study aimed to study the specific molecular regulatory mechanism of baicalein in anti-osteosarcoma and the possible regulatory signaling network involved. In vitro experiment, MG-63 cells treated with 0, 50, 75, and 100 μM of baicalein. Cell viability, proliferation, migration, invasion, cycle, apoptosis, and morphology were detected using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazoliumbromide (MTT), clone formation, wound healing, Transwell, flow cytometry, Hoechst staining, wound healing and microscopic observation. In addition, cycle, apoptosis and EMT-related proteins and p-ERK/ERK expression level were analyzed using western blotting. In vivo experiments were performed by BALB/c-nude mice model establishment to detect mice and tumor weight, tumor volume, positive rate and p-ERK/ERK expression when mice treated with 100 μM of baicalein. Firstly, the IC of baicalein was 67.57 μM. Then, baicalein decreased cell viability, proliferation, migration, invasion, and the expression of CDK2, Cyclin D1, Cyclin E1, Bcl-2, N-cad, Vimentin, MMP-2, MMP-9, p-ERK/ERK, while increased G1 phase numbers, apoptosis and the expression level of p21, p27, cleaved caspase 3/9, Bax, E-cad, ZO-1 in a dose-dependent manner in MG-63 cells. Also, baicalein reduced the body weight, tumor weight and volume and relative expression level of p-ERK/ERK in vivo. Baicalein inhibits cell development, metastasis, and EMT progress and induces cell cycle arrest and apoptosis by regulating ERK signaling pathway in osteosarcoma, and has a visible anti-osteosarcoma effect in vivo.
骨肉瘤是一种高度恶性的原发性肿瘤。黄芩苷具有广谱抗肿瘤作用。本研究旨在探讨黄芩苷抗骨肉瘤的具体分子调控机制及可能涉及的调控信号网络。体外实验中，用0、50、75和100μM的黄芩苷处理MG-63细胞。采用3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四氮唑溴盐(MTT)、克隆形成、伤口愈合、Transwell、流式细胞术、Hoechst染色、伤口愈合及显微镜观察等方法检测细胞活力、增殖、迁移、侵袭、周期、凋亡及形态。此外，采用蛋白质免疫印迹法分析周期蛋白、凋亡相关蛋白及上皮-间质转化(EMT)相关蛋白和p-ERK/ERK表达水平。体内实验通过建立BALB/c裸鼠模型，检测给予100μM黄芩苷处理后小鼠的体重、肿瘤重量、肿瘤体积、阳性率及p-ERK/ERK表达。首先，黄芩苷的半数抑制浓度(IC)为67.57μM。然后，黄芩苷以剂量依赖的方式降低MG-63细胞的活力、增殖、迁移、侵袭以及细胞周期蛋白依赖性激酶2(CDK2)、细胞周期蛋白D1(Cyclin D1)、细胞周期蛋白E1(Cyclin E1)、B细胞淋巴瘤-2(Bcl-2)、神经钙黏蛋白(N-cad)、波形蛋白(Vimentin)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、p-ERK/ERK的表达，同时增加G1期细胞数量、凋亡以及p21、p27、裂解的半胱天冬酶3/9、Bax、E-钙黏蛋白(E-cad)、紧密连接蛋白1(ZO-1)的表达水平。此外，黄芩苷在体内降低了小鼠体重、肿瘤重量和体积以及p-ERK/ERK的相对表达水平。黄芩苷通过调节骨肉瘤中的ERK信号通路抑制细胞发育、转移和EMT进程，并诱导细胞周期阻滞和凋亡，在体内具有明显的抗骨肉瘤作用。
