维替泊芬影响骨肉瘤MG63细胞增殖和迁移侵袭的作用机制  

The functional mechanism of veterporfin affecting the proliferation,migration and invasion of osteosarcoma MG-63 cell line

作　　者：陈蔚[1] 余铃[2] 陈敬腾 夏露[1] 郭卫春[2] CHEN Wei;YU Ling;CHEN Jingteng;XIA Lu;GUO Weichun(Department 3 of Orthopaedics,Renmin Hospital of Wuhan University,Wuhan,Hubei 430060,China;Department 1 of Orthopaedics,Renmin Hospital of Wuhan University,Wuhan,Hubei 430060,China)

机构地区：[1]武汉大学人民医院骨3科,湖北武汉430060 [2]武汉大学人民医院骨1科,湖北武汉430060

出　　处：《安徽医药》2019年第12期2337-2341,共5页Anhui Medical and Pharmaceutical Journal

基　　金：国家自然科学基金青年科学基金项目(81502575)

摘　　要：目的探讨维替泊芬影响骨肉瘤MG63细胞增殖及迁移侵袭的的作用机制。方法体外培养骨肉瘤MG63细胞,采用人胆囊收缩素/缩胆囊素八肽(CCK 8)检测不同浓度(0、2、4、6、8、10μmol/L)维替泊芬对骨肉瘤MG63细胞增殖影响;流式细胞术分析(0、2、4、6、8μmol/L)维替泊芬对骨肉瘤MG63细胞周期分布及凋亡的影响;划痕实验和TranswellTM侵袭实验检测(0、2、4、6、8μmol/L)维替泊芬对骨肉瘤MG63细胞迁移及侵袭能力的影响;蛋白质印迹法(Western Blot)检测(0、2、4、6、8μmol/L)维替泊芬对Hippo信号通路中Yes 相关蛋白1(YAP1)、TEA转录因子1(TEAD1)、磷酸化Yes 相关蛋白1(pYAP1)的蛋白表达水平。结果CCK 8结果显示维替泊芬能够抑制骨肉瘤MG63细胞的增殖,24、48、72 h的半抑制浓度(IC50)分别为(6.592±0.121)μmol/L、(4.668±0.075)μmol/L、(2.953±0.078)μmol/L,并呈时间、浓度依赖性;流式细胞术结果表明维替泊芬以浓度依赖的方式诱导骨肉瘤MG63细胞凋亡,使骨肉瘤MG63细胞周期阻滞于G0/G1期;划痕实验和TranswellTM侵袭实验结果表明维替泊芬可抑制MG63细胞迁移和侵袭;蛋白质印迹法结果显示维替泊芬处理的MG63细胞中YAP1、TEAD1蛋白表达水平降低,而pYAP1蛋白表达水平保持不变。结论维替泊芬能抑制骨肉瘤MG63细胞增殖、促进细胞凋亡、导致细胞周期阻滞,以及抑制细胞迁移和侵袭,并且下调YAP1及TEAD1表达,维替泊芬阻碍YAP1和TEAD1相互作用可能是抗骨肉瘤的作用机制。Objective To explore the functional mechanism of veterporfin on proliferation,migration and invasion of osteosarcoma MG 63 cell line.Methods The osteosarcoma MG 63 cells were cultured in vitro.Human cholecystokinin octapeptide(CCK 8)as say was used to detect the anti proliferative effects of(0,2,4,6,8,10μmol/L)veterporfin on osteosarcoma MG 63 cells.The ef fects of(0,2,4,6,8μmol/L)veterporfin on cell cycle and apoptosis were analyzed by flow cytometry.The wound healing assay and TranswellTM invasion assay were used to determine the effects of(0,2,4,6,8μmol/L)veterporfin on cell migration and invasion ca pability.The effects of(0,2,4,6,8μmol/L)veterporfin on the expression levels of Yes associated protein 1(YAP1),TEA domain family member 1(TEAD1)and phosphorylate Yes associated protein 1(pYAP1)in Hippo signaling pathway were determined by Western blotting.Results CCK 8 assay results showed that veterporfin inhibited the proliferation of MG 63 cells.The semi inhibitory concentrations(IC50)at 24,48,and 72 h were(6.592±0.121)μmol/L,(4.668±0.075)μmol/L,and(2.953±0.078)μmol/L,re spectively,which were time and concentration dependent.Flow cytometry showed that veterporfin could cause osteosarcoma cell cy cle arrest in G0/G1 phase and induce apoptosis of osteosarcoma MG 63 cells in a concentration dependent manner.Wound healing assay and TranswellTM invasion assay confirmed that veterporfin could inhibit the migration and invasion ability of MG 63 cells.The Western blot assay indicated that the expressions of YAP1,TEAD1 were reduced,whereas the expression of pYAP1 kept un changed.Conclusions Veterporfin inhibited the proliferation of osteosarcoma cells,promoted cell apoptosis,led to cell arrest,inhib ited cell migration and invasion,and downregulated the expression levels of YAP1 and TEAD1.Veterporfin blocking the interaction of YAP1 and TEAD1 might be the mechanism of targeting osteosarcoma.

关 键 词：骨肉瘤 维替泊芬 Hippo信号通路 增殖 迁移 侵袭 流式细胞术 细胞迁移分析 印迹法 蛋白质 人胆囊收缩素/缩胆囊素八肽 

分 类 号：R73[医药卫生—肿瘤]