| 吴良邦,葛云林.薯蓣皂苷通过调控AMPK/mTOR自噬信号通路影响骨肉瘤细胞增殖及侵袭的机制研究[J].浙江中西医结合杂志,2024,34(4): |
| 薯蓣皂苷通过调控AMPK/mTOR自噬信号通路影响骨肉瘤细胞增殖及侵袭的机制研究 |
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| 投稿时间：2023-06-14 修订日期：2024-04-14 |
| DOI： |
| 中文关键词: 薯蓣皂苷 骨肉瘤 自噬 AMPK/mTOR信号通路 |
| 英文关键词:Dioscin osteosarcoma autophagy AMPK/mTOR pathway |
| 基金项目:浙江省教育厅一般科研项目,编号:Y202249274 |
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| 中文摘要: |
| 【】 目的 探讨薯蓣皂苷(Dioscin)通过调控AMPK/mTOR自噬信号通路对MG63细胞增殖及侵袭的影响。方法 体外培养骨肉瘤MG-63细胞,分为空白对照组、薯蓣皂苷不同剂量组(20μmol/L、40μmol/L、80μmol/L、100μmol/L),CCK-8法观察细胞增殖能力,划痕实验观察细胞迁移能力,通过Transwell小室观察细胞侵袭能力,丹酰尸胺法(MDC)观察细胞自噬情况,Western blot检测磷酸化腺苷酸活化蛋白激酶/腺苷酸活化蛋白激酶(p-AMPK/AMPK)、磷酸化哺乳动物雷帕霉素靶蛋白/哺乳动物雷帕霉素靶蛋白(p-mTOR/mTOR)、转录因子EB(TFEB)、被微管相关蛋白轻链3II(LC3-Ⅱ)、肌球蛋白样BCL2结合蛋白(beclin-1)蛋白表达水平。结果 与空白对照组比较,Dioscin各剂量组MG63细胞增殖率、细胞迁移及侵袭能力依次降低,呈浓度依赖性(P＜0.05) ; MDC染色显示,空白对照组中仅有少量细胞自噬泡,作用48h后Dioscin各组自噬泡依次增多；与空白对照组相比,Dioscin各组MG63细胞中p-AMPK/AMPK、TFEB、LC3-Ⅱ、beclin-1蛋白水平依次升高,p-mTOR/mTOR蛋白水平依次降低,差异均具有统计学意义(P＜0.05)；与Dioscin组比较,Dioscin+AMPK抑制剂组p-AMPK/AMPK、TFEB、LC3-Ⅱ、beclin-1蛋白表达水平明显降低,p-mTOR /mTOR表达升高,差异均具有统计学意义(P＜0.05)。结论 Dioscin可能通过调控AMPK/mTOR-TFEB通路抑制骨肉瘤细胞增殖、迁移及侵袭。 |
| 英文摘要: |
| 【】 Objective: To explore the effect of Dioscin on the proliferation and invasion of MG63 cells by regulating the AMPK/mTOR autophagy signaling pathway. Methods: Osteosarcoma MG-63 cells were cultured in vitro and divided into blank control group and Dioscin groups with different doses (20μmol/L、40μmol/L、80μmol/L、100μmol/L). Determinations of cell proliferation by CCK-8 method,cell migration ability by scratch experiment,cell invasion ability by Transwell, observation of autophagy by danylcadaverine method(MDC),the protein expression levels of p-AMPK/AMPK,p-mTOR/mTOR,TFEB,LC3-Ⅱ,beclin-1 by Western blot.Results: Compared with the blank control group, the proliferation rate, cell migration and invasion ability of MG63 cells in each dose group of Dioscin decreased sequentially in a concentration-dependent manner (P＜0.05); MDC staining showed that there were only a small number of autophagic vesicles in the blank control group, after 48 hours of exposure,the number of autophagic vesicles in each Dioscin group increased sequentially; compared with the blank control group, the protein levels of p-AMPK/AMPK,TFEB,LC3-Ⅱ,and beclin-1 in MG63 cells in each Dioscin group increased,and the protein levels of p-mTOR/mTOR increased in concentration-dependent,the difference were statistically significant (P＜0.05)；compared with Dioscin group, p-AMPK/AMPK, TFEB, LC3-Ⅱ, beclin-1 decreased in Dioscin+AMPK inhibitors group,p-mTOR/mTOR increases, the differences were statistically significant(P<0.05). |
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