二甲双胍对胰腺癌和胰腺癌干细胞中ALDH1表达的作用

发布时间：2018-09-07 21:28

【摘要】：目的观察二甲双胍对胰腺癌PANC-1细胞和胰腺癌干细胞中ALDH1表达的影响,探讨其作用机制。方法采用CCK-8和Western blot检测不同浓度二甲双胍对胰腺癌PANC-1细胞的增殖及ALDH1和p-mTOR蛋白表达的影响。Western blot、免疫荧光法及q RT-PCR检测二甲双胍、雷帕霉素以及联合用药组PANC-1细胞中ALDH1、4EBP1、p-m TOR蛋白及m RNA的表达。超低黏附培养板悬浮培养胰腺癌细胞后观察干细胞成球数目的改变以及Western blot检测干细胞ALDH1的表达;建立胰腺癌裸鼠移植瘤模型比较PANC-1细胞以及胰腺癌成球细胞的成瘤能力。结果二甲双胍抑制胰腺癌PANC-1细胞的增殖,且呈时间剂量依赖性,最佳干预时间为48 h,IC50约为20 mmol/L;与对照组相比,二甲双胍、雷帕霉素和联合用药组ALDH1、4EBP1、p-mTOR的ALDH1的蛋白表达以及m RNA水平明显降低(P0.05)。胰腺癌成球细胞的体外成瘤能力以及ALDH1的表达比PANC-1细胞组显著增高;相比于对照组,二甲双胍、雷帕霉素和联合用药组胰腺癌细胞的成球能力以及胰腺癌干细胞中ALDH1的表达均显著降低(P0.05)。结论二甲双胍以及雷帕霉素均能显著抑制胰腺癌PANC-1细胞和胰腺癌干细胞ALDH1的表达,其作用机制可能是通过抑制mTOR通路。
[Abstract]:Aim to investigate the effect of metformin on the expression of ALDH1 in pancreatic cancer PANC-1 cells and pancreatic cancer stem cells. Methods CCK-8 and Western blot were used to detect the effect of metformin at different concentrations on the proliferation of PANC-1 cells and the expression of ALDH1 and p-mTOR. Western blot, immunofluorescence assay and Q RT-PCR were used to detect metformin. The expression of ALDH1,4EBP1,p-m TOR protein and m RNA in PANC-1 cells of rapamycin group and combined treatment group. After suspension culture of pancreatic cancer cells with ultra-low adhesion culture plate, the number of stem cells and the expression of ALDH1 of stem cells were detected by Western blot, and the tumorigenic ability of PANC-1 cells and pancreatic cancer ball cells were compared by establishing nude mouse xenograft tumor model of pancreatic cancer. Results Metformin inhibited the proliferation of pancreatic cancer PANC-1 cells in a dose-dependent manner. The best intervention time was 48 h and IC50 was about 20 mmol/L;. The expression of ALDH1 protein and the level of m RNA in ALDH1,4EBP1,p-mTOR were significantly decreased in rapamycin and combination groups (P0.05). The tumor forming ability and ALDH1 expression of pancreatic cancer ball cells were significantly higher than those of PANC-1 cells, and the expression of metformin was significantly higher than that of the control group. The colonization ability of pancreatic cancer cells and the expression of ALDH1 in pancreatic cancer stem cells were significantly decreased in rapamycin and combination groups (P0.05). Conclusion Metformin and rapamycin can significantly inhibit the expression of ALDH1 in pancreatic cancer PANC-1 cells and pancreatic cancer stem cells by inhibiting the mTOR pathway.
【作者单位】：郑州大学第一附属医院肿瘤科;
【分类号】：R735.9

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