摘要
目的 分析二甲双胍激活Hippo信号通路对胰腺癌细胞生长的抑制作用。方法 采用中国科学院上海生命科学院细胞资源中心提供的人胰腺癌细胞株PANC-1,进行细胞培养,提取细胞Total RNA,测定RNA样本纯度与浓度,RNA逆转录为cDNA。实时荧光定量Real-time PCR提取细胞总蛋白,采用BCA染色法测定蛋白浓度并进行蛋白变性,蛋白质印迹法检测目的蛋白表达,分析蛋白RNA样品质检情况。二甲双胍组采用二甲双胍处理,对照组不采用二甲双胍处理,统计分析二甲双胍对人约克同源(YAP1)、大肿瘤抑制基因1(LATS1)、育系20样激酶1(MST1)mRNA表达的调节作用,并统计分析二甲双胍对P-YAP1、LATS1、MST1蛋白质表达的影响。结果 所有样品质检均合格,能够进行后续检测。二甲双胍组PANC-1细胞中YAP1、LATS1、MST1 mRNA表达分别为4.16±0.36、3.72±0.85、4.04±0.47,均明显高于对照组(1.02±0.10、0.94±0.04、1.01±0.08),差异均有统计学意义(P<0.05)。二甲双胍组PANC-1细胞中P-YAP1、LATS1、MST1蛋白质表达分别为0.66±0.01、0.73±0.01、0.50±0.01,均明显高于对照组(0.16±0.01、0.14±0.01、0.32±0.01),差异均有统计学意义(P<0.05),但两组YAP1蛋白质表达之间的差异无统计学意义(P>0.05)。结论 二甲双胍治疗胰腺癌能够激活Hippo信号通路,从而抑制细胞生长,发挥抗肿瘤作用。
Objective To analyze the inhibitory effect of metformin on pancreatic cancer cell growth by activating Hippo signaling pathway.Methods Human pancreatic cancer cell line PANC-1 provided by Cell Resource Center of Shanghai Academy of Biological Sciences,Chinese Academy of Sciences was purchased for cell culture.Total RNA was extracted,the sample purity and concentration of RNA was determined,RNA was reverse-transcribed into cDNA,and total protein was extracted by Real-time fluorescent quantitative real-time PCR.The protein concentration was determined by BCA colorimetry and the protein denaturation was did,Western blotting was used to detect the target protein expression.The metformin group was treated with metformin,while the control group did not receive metformin.Statistical analysis was conducted on the regulatory effects of metformin on the mRNA expression of human York homology(YAP1),large tumor suppressor gene 1(LATS1),and germline 20 like kinase 1(MST1).The effects of metformin on P-YAP1,LATS1 and MST1 protein expression were analyzed.Results All samples were qualified for follow-up testing.The expressions of YAP1,LATS1 and MST1 mRNA in PANC-1 cells in metformin group were 4.16±0.36,3.72±0.85 and 4.04±0.47,respectively,which were significantly higher than those in control group(1.02±0.10,0.94±0.04 and 1.01±0.08),and the differences were statistically significant(P<0.05).The protein expressions of P-YAP1,LATS1 and MST1 in PANC-1 cells in metformin group were 0.66±0.01,0.73±0.01 and 0.50±0.01,respectively,which were significantly higher than those in control group(0.16±0.01,0.14±0.01 and 0.32±0.01),and the differences were statistically significant(P<0.05),but there was no statistically significant difference in YAP1 protein expression between the two groups(P>0.05).Conclusion Metformin in the treatment of pancreatic cancer can activate the Hippo signaling pathway,thus inhibiting cell growth and bringing out the anti-tumor effect.
作者
李世艳
孙永辉
余鸿
LI Shi-yan;SUN Yong-hui;YU Hong(Department of Pancreatic Surgery,First Affiliated Hospital of Xinjiang Medical University,Urumqi Xinjiang 830054,China)
出处
《临床和实验医学杂志》
2024年第18期1905-1908,共4页
Journal of Clinical and Experimental Medicine
基金
新疆维吾尔自治区自然科学基金项目(编号:SYTG-Y202321)。
