基于ERK1/2通路探讨八宝丹抑制骨肉瘤生长的机制  

Mechanism of Babaodan Inhibiting Growth of Osteosarcoma via ERK1/2 Signaling Pathway

作　　者：陈鹏 张冬龙[3] 刘宇 林庆宾 刘俊宁[1,2] 洪振强 CHEN Peng;ZHANG Donglong;LIU Yu;LIN Qingbin;LIU Junning;HONG Zhenqiang(College of Traditional Chinese Medicine,Fujian University of Traditional Chinese Medicine,Fujian Fuzhou 350122,China;Key Laboratory of Bone Injury and Sports Rehabilitation of Traditional Chinese Medicine,Ministry of Education,Fujian Fuzhou 350122,China;Xiamen Jimei Street Community Health Service Center,Fujian Xiamen 361021,China)

机构地区：[1]福建中医药大学中医学院,福建福州350122 [2]中医骨伤及运动康复教育部重点实验室,福建福州350122 [3]厦门市集美街道社区卫生服务中心,福建厦门361021

出　　处：《福建中医药》2023年第6期18-21,51,共5页Fujian Journal of Traditional Chinese Medicine

基　　金：福建省自然科学基金项目(2019J01726)。

摘　　要：目的通过观察八宝丹(BBD)对U2OS细胞生长的抑制作用以及对细胞外调节蛋白激酶(ERK1/2)通路的影响,以期揭示八宝丹抑制骨肉瘤生长的机制。方法选用U2OS细胞株作为研究对象,实验分为对照组和八宝丹高、中、低剂量组,高剂量组药物浓度为1.5 mg/mL;中剂量组药物浓度为1.0 mg/mL、低剂量组药物浓度为0.5 mg/mL,干预结束后采用CCK8法检测八宝丹对U2OS细胞增殖的影响;流式细胞术检测八宝丹对细胞凋亡的影响;qPCR检测八宝丹对ERK1/2通路上游Ras、Raf基因表达水平的影响;Western blot检测八宝丹对Cy⁃clinD1、MEK1/2、ERK1/2和p-ERK1/2蛋白表达量的影响。结果与对照组比较,不同浓度的八宝丹在干预24 h和48 h对U2OS细胞均有不同程度的生长抑制作用(P均<0.05);不同浓度八宝丹干预48 h均能明显降低Ras、Raf基因的表达,同时下调CyclinD1、MEK1/2蛋白的表达,下调ERK1/2与p-ERK1/2蛋白比率(P均<0.05)。结论八宝丹可抑制U2OS骨肉瘤细胞增殖,促进其凋亡,可能是通过抑制ERK1/2信号通路实现的。Objective:To reveal the mechanism of Babaodan(BBD)inhibiting the growth of osteosarcoma,and provide a basis for the promotion of its clinical efficacy against osteosarcoma by observing the effects of Babaodan on the growth of U2OS osteosarcoma cells via the extracellular regulatory protein kinase(ERK1/2)signaling pathway.Methods:Osteosarcoma cell lines of U2OS were selected as the study subjects,and the experiment was divided into control group and Babaodan groups with different concentrations(BBD-0.5 mg/mL group,BBD-1 mg/mL group,BBD-1.5 mg/mL group).After the intervention,CCK8 method was used to detect the effect of BBD on the proliferation of U2OS cells.Flow cytometry was used to detect the effect of BBD on cell apoptosis.qPCR was used to detect the effect of BBD on the expression of Ras and Raf genes in the upstream of ERK1/2 pathway.The effects of BBD on the expression of CyclinD1,MEK1/2,ERK1/2 and p-ERK1/2 proteins were detected by Western blot analysis.Results:Compared with the control group,different concentrations of BBD inhibited the growth of U2OS cells at 24 h and 48 h after intervention(P<0.05).BBD intervention for 48 h could significantly reduce the expressions of Ras and Raf genes,down-regulate the expressions of CyclinD1 and MEK1/2 proteins,and the ratio of ERK1/2 to P-erk1/2 proteins(P<0.05).Conclusion:BBD can inhibit the proliferation of U2OS osteosarcoma cells and promote their apoptosis,which may be achieved by inhibiting ERK1/2 signaling pathway.

关 键 词：骨肉瘤 八宝丹 U2OS细胞 增殖 凋亡 ERK1/2信号通路 

分 类 号：R285[医药卫生—中药学]