Abstract: Objective: To investigate the inhibitory effect of metformin on human oral squamous cell carcinoma SCC-4 and CAL-27 cells. Methods: The recovery, culture and subculture of oral squamous cell carcinoma SCC-4 cell line and CAL-27 cell line were performed. The effects of metformin on the proliferation, cell activity, clone formation and growth cycle of human oral squamous cell carcinoma SCC-4 and CAL-27 cells were examined by MTT assay, flow cytometry and Western blot. Results: SCC-4 and CAL-27 were treated with metformin at 5 mmol/L for 12 h, 24 h and 48 h respectively, the proportion of cells in G0/G1 phase increased in a time-dependent manner (P<0.05), but the proportion of cells in S phase and G2/M phase decreased (P<0.05). The cell viability of SCC-4 and CAL-27 was significantly decreased and showed in a dose-dependent manner after 48 h of metformin intervention. After treated with metformin for 2 weeks, clone formation of SCC-4 and CAL-27 decreased in a dose-dependent manner (P<0.05). After SCC-4 and CAL-27 were treatedwith 5 mmol/L metformin, the level of AMPKa (Thrl72) was significantly increased. The expression of mTOR and its downstream direct acting molecules  p-mTOR (Ser2448), p-S6Kl (Thr389) and p-4E-BPl (Thr37 / 46) were significantly decreased in a time-dependent manner. Conclusion: Metformin through the activating of AMPK signaling pathway to inhibits mTOR signaling pathway to suppress the cell viability, cell proliferation and clone formation of SCC-4 and CAL-27 cells.

Key words: metformin, oral squamous cell carcinomas, SCC-4 cells, CAL-27 cells, molecular mechanisms

摘要： 目的：探讨二甲双胍对于人口腔鳞癌SCC-4和CAL-27细胞的抑制作用。方法：对口腔鳞癌SCC-4细胞株和CAL-27细胞株进行复苏、培养和传代处理，运用MTT检测法、流式细胞术检测法、Western blot检测法，观察二甲双胍对人口腔鳞癌SCC-4和CAL-27细胞的体外增殖能力、细胞活性、克隆形成以及生长周期的影响。结果：SCC-4和CAL-27在二甲双胍干预48 h后细胞活性降低，且呈现明显的浓度依赖性；SCC-4和CAL-27在5 mmol/L浓度的二甲双胍干预12、24、48 h后，G0/G1期所占比例随时间推移而逐渐增高（P<0.05），但S期和G2/M期所占的比例却减小（P<0.05）；SCC-4和CAL-27经二甲双胍处理2周后，细胞的克隆数显著减少，且呈现明显的浓度依赖性（P<0.05）；SCC-4和CAL-27经5 mmol/L二甲双胍干预后，AMPKa憐酸化水平（Thrl72）显著提升，mTOR及其下游直接作用分子p-mTOR (Ser2448)、p-S6Kl (Thr389)、p-4E-BPl (Thr37/46)表达量显著下降，且有时间依赖性。结论：二甲双胍主要通过激活AMPK信号通路而抑制mTOR信号通路，来抑制口腔鳞癌SCC-4和CAL-27细胞的细胞活性，细胞增殖和克隆形成，使口腔鳞癌SCC-4和CAL-27细胞生长阻滞在G0/G1期，从而达到治疗口腔鳞癌的目的。

关键词: 二甲双胍；  , 口腔鳞癌；  , SCC-4细胞；  , CAL-27细胞；  , 分子机制

CLC Number: 

• R782

• R739.8