荜茇酰胺通过诱导口腔鳞癌细胞自噬抑制增殖  

Piperlongumine Suppresses Proliferation of Oral Squamous Cell Carcinoma via Promoting Autophagy

作　　者：许小鸿 郅程[2] 袁忠民[3] XU Xiao-hong;ZHI Cheng;YUAN Zhong-min(Department of Stomatology,The Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,China;Department of Pathology,The Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,China;Institute of Neuroscience,The Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,China)

机构地区：[1]广州医科大学附属第二医院口腔科,广东广州510260 [2]广州医科大学附属第二医院病理科,广东广州510260 [3]广州医科大学附属第二医院神经科学研究所,广东广州510260

出　　处：《中山大学学报（医学科学版）》2021年第4期543-549,共7页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：广东省中医药局基金(20181158);广州市科学技术局基金(202002020088)。

摘　　要：【目的】探讨荜茇酰胺(PPLGM)对口腔癌存活的影响及机制。【方法】体外培养口腔鳞状细胞癌Cal27及UM1细胞株,经不同浓度的荜茇酰胺处理后,MTT比色法检测荜茇酰胺对增殖的影响,蛋白质印迹法检测胞内微管相关蛋白轻链3-I、3-Ⅱ(LC3-I、LC3-Ⅱ)、Beclin1、p62、雷帕霉素靶蛋白(mTOR)及ribosomal S6(S6)的表达或活性变化;用自噬抑制剂3-Methyladenine(3-MA)或者Bafilomycin A1(BAFA1)和荜茇酰胺共处理细胞后,检测LC3-Ⅱ、p62表达和细胞活力。【结果】与对照组比较,荜茇酰胺显著抑制Cal27及UM1细胞株增殖能力,呈浓度及时间依赖性(P<0.05);Western blot结果显示,不同浓度(1.0、3.0和5.0μmol/L)荜茇酰胺处理细胞均使LC-I、LC3-Ⅱ表达水平显著高于对照组(P<0.05),而Beclin1和p62表达低于对照组(P<0.05);荜茇酰胺处理组中p-mTOR和p-S6磷酸化水平显著低于对照组(P<0.05);与单独荜茇酰胺处理比较,3-MA和荜茇酰胺共处理使胞内LC3-I和LC3-Ⅱ表达水平明显下降,p62表达升高(P<0.05),但BAFA1和荜茇酰胺共处理使细胞内p62表达升高(P<0.05),LC3-Ⅱ表达水平无明显改变;3-MA和B AFA1处理都能拮抗荜茇酰胺对细胞活力的抑制效应。【结论】荜茇酰胺通过抑制mTOR/S6通路活性诱导口腔鳞癌细胞发生自噬,进而抑制增殖。【Objective】To explore the effect of piperlongumine(PPLGM)on the proliferation of oral squamous cell carcinoma(OSCC)cells and the potential mechanism involved【.Methods】OSCC cell lines Cal-27 and UM1 cultured in vitro were treated with PPLGM at different doses and time courses.Then the viability of Cal-27 and UM1 cells was evaluated by MTT assay and the expression of LC3-Ⅰ,LC3-Ⅱ,Beclin1,p62 and the phosphorylation of mTOR and S6K were determined by Western blot.Autophagy inhibitors 3-Methyladenine(3-MA)at 100 nmol/L and Bafilomycin A1(BAFA1)at 400 nmol/L were administrated to observe the rescued effects on the PPLGM-treated cells.【Results】PPLGM treatment caused a dose-and time-dependent inhibition on the viability of both Cal-27 and UM1 cells(P<0.05).Western blot results showed the expression levels of LC3-Ⅰ and LC3-Ⅱ in PPLGM(1.0,3.0,5.0μmol/L)group were significantly high-er(P<0.05)and p62 expression levels were lower(P<0.05)compared with the control group.PPLGM treatment at 3.0 or 5.0μmol/L caused a significant decrease of p-mTOR and p-S6(P<0.05).Autophagy inhibitors 3-MA significantly rescued PPLGM-induced increase of LC3-I,LC-3Ⅱ and decrease of p62,while BAFA1 just recovered PPLGM-caused decrease of p62.Both 3-MA and BAFA1 could effectively rescue PPLGM-mediated inhibitory effects on the viability of cells.【Conclusion】PPLGM suppresses the viability of OSCC cells through promoting mTOR activity loss-dependent autophagy.

关 键 词：荜茇酰胺 口腔鳞状细胞癌 自噬 LC-3 MTOR 

分 类 号：R739.85[医药卫生—肿瘤]