Hippo-YAP信号通路活化参与二甲双胍抑制胰腺癌细胞生长的作用  

Hippo-YAP signaling pathway activation was involved in the inhibitory effect of mefformin on the growth of pancreatic cancer PANC1 cells

作　　者：李映璇 高绥之 苏松[1] 徐茂锦[1] 高军[2] 诸娴[2] 金晶[2] 吴洪玉[2] 

机构地区：[1]第二军医大学长海医院内分泌科,上海200433 [2]第二军医大学长海医院消化内科,上海200433 [3]第二军医大学学员旅

出　　处：《中华胰腺病杂志》2016年第2期82-86,共5页Chinese Journal of Pancreatology

基　　金：国家自然科学基金（81272663,81472279）;上海市教育委员会科研创新基金（13ZZ062）

摘　　要：目的观察二甲双胍对人胰腺癌PANC1细胞增殖、细胞周期及凋亡的影响，探讨其与Hippo-YAP信号通路活化的关系。方法应用5、10、20、40 mmol/L的二甲双胍分别处理人胰腺癌PANC1细胞24、48、60、72 h，以不加二甲双胍处理的细胞作为对照。采用CCK-8法检测细胞增殖，流式细胞术检测细胞周期和细胞凋亡，实时荧光定量PCR法检测细胞YAP mRNA表达，蛋白质印迹法检测细胞YAP1、磷酸化YAP1（p-YAP1）蛋白表达。结果二甲双胍呈剂量依赖性抑制PANC1细胞的增殖，各组间差异均有统计学意义（P值均〈0.01）。二甲双胍抑制PANC1细胞增殖的IC50值为20 mmol/L。用20 mmol/L二甲双胍处理细胞48 h后细胞的G1期、S期细胞百分比及YAP1 mRNA表达量、p-YAP1蛋白表达量分别为（77.12±1.22）%、（9.13±0.73）%、4.17±0.37、0.67±0.01；对照组分别为（60.75±1.53）%、（26.97±1.18）%、1.03±0.11、0.17±0.01，两组间的差异均有统计学意义（P值均〈0.01）。对照组与二甲双胍处理组细胞的凋亡率分别为（5.65±1.19）%、（9.83±1.36）%，YAP1蛋白表达量分别为0.42±0.00、0.41±0.00，两组的差异均无统计学意义（P值均﹥0.05）。结论二甲双胍可显著抑制人胰腺癌PANC1细胞的增殖，使细胞周期阻滞在G0/G1期，其机制可能与Hippo-YAP信号通路活化有关。Objective To investigate the effects of metformin on the proliferation, cell cycle and apoptosis in human pancreatic cancer PANC1 cells, and to explore their relations with Hippo-YAP signaling pathway activation. Methods Pancreatic cancer PANC1 cells were treated with 5, 10, 20, 40 mmol/L mefformin for 24 h, 48 h, 60 h, 72 h, respectively. Meanwhile, the PANC1 cells that were not treated with metformin were set as the controls. CCK-8 method was used to examine the proliferation of PANC1 cells. Flow cytometry was used to detect cell cycle and apoptosis, and YAP1 mRNA was detected by real-time fluorescent quantitative PCR, and YAP1 and pYAP1 protein were detected by Western blot, respectively. Results Treatment with metformin could observably inhibit the proliferation of PANC1 cells in a dose-dependent manner, and the difference among different groups was statistically significant （all P〈0.01 ）. The half inhibition concentration （ IC50 ） of mefformin was about 20 mmol/L. After treated with 20 mmol/L mefformin for 48 h, the proportion of G1 cells and S phase cells, the expression of YAP1 mRNA and p-YAP1 protein was （77.12 ± 1.22）% ,（9.13 ±0.73）% ,4.17 ±0.37,0.67 ±0.01, while in control group, these were （60.75 ± 1.53）%, （ 26.97 ± 1.18 ）%, 1.03 ± 0. 11,0. 17 ± 0.01, and the difference was statistically significant （ P 〈 0.01 ）. The apoptosis rate was （ 5.65 ± 1.19） % vs （ 9.83 ± 1.36） % and YAP1 protein expression was （0.42 ±0.00） vs （0.41 ±0.00）. The differences were not statistically significant（all P 〉 0.05）. Conclusions Mefformin could significantly inhibit the proliferation of pancreatic cancer PANC1 cells and lead to G0/G1 phase arrest, in which the activation of Hippo-YAP signaling pathway may be involved.

关 键 词：胰腺肿瘤 二甲双胍 细胞增殖 细胞凋亡 YAP 信号通路 

分 类 号：R735.9[医药卫生—肿瘤]