抑制细胞自噬显著增加二甲双胍诱导的口腔癌细胞凋亡  

Inhibited autophagy enhance metformin induced apoptosis in oral squamous cell carcinoma

作　　者：朱非亚[1] 陈林[1] 皮会丰[2] 王铠[1] 

机构地区：[2]中南大学湘雅二医院口腔医学中心,湖南长沙410011 [3]第三军医大学预防医学院劳动卫生学教研室,重庆400038

出　　处：《局解手术学杂志》2016年第10期703-707,共5页Journal of Regional Anatomy and Operative Surgery

基　　金：国家自然科学基金(81500832);湖南省自然科学基金(2012Fj6119)

摘　　要：目的研究二甲双胍对人口腔癌细胞增殖及凋亡的影响,为口腔癌的治疗开辟新的路径。方法采用不同浓度(5、10、20 mmol/L)二甲双胍处理CAL27细胞24、48、72 h;流式细胞术和TUNEL检测细胞凋亡数;免疫荧光观察自噬小体;Western blot法检测凋亡和自噬标志物的表达。结果不同浓度二甲双胍处理CAL27细胞可显著诱导其凋亡;CL-PARP、Bcl-2等凋亡标志物也显著增高。二甲双胍可以刺激CAL27细胞产生自噬,细胞荧光检测到增多的GFP-LC3小点;LC3、Beclin1等自噬标志蛋白也显著变化。二甲双胍诱导CAL27细胞STAT3表达下调的同时可以抑制mTOR信号通路是其产生自噬的可能原因。抑制自噬能够显著加强二甲双胍诱导的肿瘤细胞凋亡。结论二甲双胍能诱导口腔癌细胞凋亡,可使口腔癌细胞自噬水平上升,其机制与STAT3及mTOR信号通路有关,抑制自噬能够显著加强二甲双胍诱导的肿瘤细胞凋亡。Objective To study the effects of mefformin on proliferation and apoptosis of oral squamous cells so as to create a new path for the treatment of oral carcinoma. Methods CAL27 cells were treated with different dosage of metformin （5,10,20 mmol/L） for 24,48, and 72 hours. The number of apoptosis was detected by flow cytometry and TUNEL. The autophagic vacuole was detected by immunofluores- cence. The expression of hallmark of apoptosis and autophagy was detected by Western blot. Results Metformin could induce apoptosis in CAL27 cells. The CL-PARP and Bcl-2 expression significantly increased. In line with the apoptosis, metformin can trigger autophagy in CAL27 cells. The expression of LC3, Beclin-1 and GFP-LC3 positive autophagosomes were increased by metformin. Mefformin inhibit the expression of STAT3 and mTOR signaling pathways at the same time might be the possible reasons of the autophagy. And inhibited autophagy could en- hance mefformin induced Caspase-3 activity in CAL27 cells. Conclusion Inactivation of STAT3 and mTOR pathway contributes to metform- in-induced autophagy. Inhibited autophagy could enhance mefformin induced apoptosis in oral squamous cell carcinoma.

关 键 词：口腔癌 二甲双胍 凋亡 自噬 MTOR STAT3 

分 类 号：R739.86[医药卫生—肿瘤]