Department of Urology, The Third Affiliated Hospital of Soochow University, No.185, Juqian Street, Tianning District, Changzhou, 213000, Jiangsu Province, China.
Department of Urology, The Affiliated Cancer Hospital of Guizhou Medical University, Guiyang, Guizhou Province, China.
BMC Urol. 2022 May 24;22(1):79. doi: 10.1186/s12894-022-01027-2.
To observe and explore the effect of metformin on the migration and proliferation of bladder cancer T24 and 5637 cells in vitro.
Bladder cancer T24 and 5637 cell lines were cultured in vitro, and were divided into group A (blank control group) and group B (metformin group: 5, 10, 15, and 20 mmol/L); both groups were plated on 6-well plates at the same time. Culture in 24-well plates was used for wound healing assays and in 96-well plates for Transwell migration and invasion, and Cell Counting Kit-8 proliferation experiments. We observed and detected the cell migration and proliferation ability of each group at 48 h, and calculated the cell migration area and survival rate. Flow cytometry was used to detect cell apoptosis in the groups. The apoptosis-related proteins, cleaved-caspase 3, cleaved-PARP, and the PI3K/AKT/mTOR signaling pathway member proteins PI3K, phosphorylated (p)-PI3K, AKT, p-AKT, mTOR, and p-mTOR were detected using western blotting.
After 48 h of treatment with different concentrations of metformin, the cell migration and proliferation capabilities were significantly lower than those in the blank control group. The proliferation and migration abilities of T24 and 5637 cells decreased in a metformin concentration-dependent manner (P < 0.05). The apoptosis rate under different concentrations of metformin, as detected by flow cytometry, showed a significantly higher rate in the metformin group than in the control group (P < 0.05). Compared with that in the control group, the level of cleaved-caspase 3 and cleaved-PARP protein in the metformin group was increased in each treatment group, and the levels of p-mTOR, p-AKT, and p-PI3K decreased significantly compared with those in the control group (P < 0.05).
Metformin inhibited bladder cancer T24 and 5637 cell migration and proliferation, and induced their apoptosis. The mechanism might involve inhibition of the activation of the PI3K/AKT/mTOR signaling pathway.
观察并探讨二甲双胍对体外膀胱癌 T24 和 5637 细胞迁移和增殖的影响。
体外培养膀胱癌 T24 和 5637 细胞系，分为 A 组（空白对照组）和 B 组（二甲双胍组：5、10、15、20mmol/L）；两组同时接种于 6 孔板，24 孔板行划痕实验，96 孔板行 Transwell 迁移及侵袭实验，细胞计数试剂盒-8 增殖实验。于 48h 观察并检测各组细胞的迁移及增殖能力，计算细胞迁移面积及存活率。流式细胞术检测各组细胞凋亡情况，Western blot 检测凋亡相关蛋白 cleaved-caspase 3、cleaved-PARP 及 PI3K/AKT/mTOR 信号通路相关蛋白 PI3K、磷酸化（p）-PI3K、AKT、p-AKT、mTOR、p-mTOR。
不同浓度二甲双胍处理 48h 后，细胞迁移及增殖能力均明显低于空白对照组，T24、5637 细胞增殖及迁移能力均呈二甲双胍浓度依赖性下降（P<0.05）；不同浓度二甲双胍作用下，流式细胞术检测细胞凋亡率显示二甲双胍组明显高于对照组（P<0.05）；与对照组相比，二甲双胍组各处理组 cleaved-caspase 3、cleaved-PARP 蛋白水平升高，p-mTOR、p-AKT、p-PI3K 水平较对照组明显降低（P<0.05）。
二甲双胍抑制膀胱癌 T24、5637 细胞迁移及增殖，诱导其凋亡，其机制可能与抑制 PI3K/AKT/mTOR 信号通路的激活有关。
