| 陈蔚,余铃,陈敬腾,等.维替泊芬影响骨肉瘤 MG63细胞增殖和迁移侵袭的作用机制[J].安徽医药,2019,23(12):2337-2341. |
| 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 |
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| DOI：10.3969/j.issn.1009?6469.2019.12.002 |
| 中文关键词: 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 |
| 英文关键词: Osteosarcoma Veterporfin Hippo signaling pathway Proliferation Migration Invasion Flow cytometry Cell migration assays Blotting,western Human cholecystokinin octapeptide |
| 基金项目:国家自然科学基金青年科学基金项目(81502575) |
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| 中文摘要: |
| 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、10 μmol/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、8 μmol/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、8 μmol/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、8 μmol/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。 |
| 英文摘要: |
| 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(0，2，4，6，8，10 μmol/L)veterporfin on osteosarcoma MG?63 cells.The ef? fects of(0，2，4，6，8 μmol/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(0，2，4，6，8 μmol/L)veterporfin on cell migration and invasion ca? pability.The effects of(0，2，4，6，8 μmol/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 healingassay and TranswellTM invasion assay confirmed that veterporfin could inhibit the migration and invasion ability of MG?63 cells.TheWestern?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 interactionof YAP1 and TEAD1 might be the mechanism of targeting osteosarcoma. |
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