没食子酸对人舌鳞状细胞癌SCC15细胞自噬的影响  

Effect of gallic acid on autophagy of human tongue squamous carcinoma cell line SCC15

作　　者：李芳[1] 关爽 LI Fang;GUAN Shuang(Stomatological Department,The Affiliated Zhongshan Hospital of Dalian University,Dalian 116001,China)

机构地区：[1]大连大学附属中山医院口腔科,辽宁大连116001

出　　处：《口腔医学》2020年第11期987-991,共5页Stomatology

基　　金：辽宁省自然基金(20180550257)。

摘　　要：目的研究没食子酸对人舌鳞状细胞癌SCC15细胞自噬的影响及其相关的分子机制。方法采用四甲基偶氮唑盐微量酶反应比色法(MTT)检测不同浓度没食子酸对SCC15细胞增殖的影响;单丹磺酰尸胺染色法(monodansylcadaverine,MDC)检测没食子酸对SCC15细胞自噬体形成的影响;应用Western blot方法检测没食子酸对SCC15细胞自噬相关蛋白LC3Ⅰ、LC3Ⅱ、Beclin1表达情况的影响,以及对PI3K/Akt/mTOR通路中p⁃Akt、p⁃mTOR蛋白表达的影响。结果MTT结果显示,90、120、150、300、600μmol/L没食子酸作用于SCC15细胞24、48 h后,SCC15细胞抑制率明显增加,且有一定的剂量依赖关系(P<0.05);MDC染色结果显示,300μmol/L没食子酸组细胞中散布于细胞质及核周的点状荧光颗粒(即自噬体)显著增多;Western blot结果显示,没食子酸作用SCC15细胞48 h后,LC3Ⅱ/LC3Ⅰ增加、Beclin 1蛋白表达上调;p⁃Akt和p⁃mTOR蛋白表达显著降低(P<0.05)。结论一定浓度没食子酸能够诱导SCC15细胞发生自噬,其作用机制可能与抑制PI3K/Akt/mTOR通路中p⁃Akt和p⁃mTOR蛋白表达,上调LC3、Beclin⁃1蛋白表达有关。Objective To observe the effects of gallic acid on autophagy of human tongue squamous cell carcinoma SCC15 cell line and investigate their related molecular mechanism.Methods MTT assay was used to detect the effect of different concentrations of gal⁃lic acid on SCC15 cell proliferation.Monodansylcadaverine(MDC)staining was used to detect the effect of gallic acid on the autopha⁃gosome formation of SCC15 cells.Western blot was applied to detect the effect of gallic acid on the protein expression of LC3Ⅰ,LC3Ⅱ,Beclin1,p⁃Akt and p⁃mTOR in PI3K/Akt/mTOR pathway.Results MTT assay showed that the inhibitory rate of SCC15 cells was significantly increased after 24 and 48 hours in the 90,120,150,300,600μmol/L gallic acid group,with a dose⁃dependent rela⁃tionship(P<0.05).MDC staining results revealed that the scattered point fluorescence particles in the cytoplasm and around the nucleus of SCC15 in the 300μmol/L gallic acid group were significantly increased.Western blot showed that gallic acid increased the ratio of LC3Ⅱ/LC3Ⅰand the protein expression of Beclin1 after 48 hours(P<0.05),while it significantly decreased the protein expressions of p⁃Akt and p⁃mTOR(P<0.05).Conclusion Gallic acid under certain concentration can induce autophagy of SCC15,whose mechanism may be related to the down⁃regulation of the protein expression of p⁃Akt and p⁃mTOR through PI3K/Akt/mTOR pathway and the up⁃regulation of LC3 and Beclin1 protein expression.

关 键 词：没食子酸 口腔鳞状细胞癌 细胞自噬 PI3K/Akt/mTOR信号通路 

分 类 号：R979.19[医药卫生—药品] R739.85[医药卫生—药学]