苯乙双胍对体内外胰腺癌血管生成的抑制作用  

The anti-angiogenesis effect of phenformin on pancreatic cancer in vitro and in vivo

作　　者：骆锴冉 刘岸 LUO Kairan;LIU An(Department of General Surgery,Affiliated Hospital of Shaoxing University,Shaoxing 312000,China;School of Chemical Engineering,Hunan Institute of Science and Technology,Yueyang 414000,China)

机构地区：[1]绍兴文理学院附属医院普外科,浙江绍兴312000 [2]湖南理工学院化工学院,湖南岳阳414000

出　　处：《健康研究》2023年第1期65-69,F0003,共6页Health Research

基　　金：湖南省卫计委科研计划项目(B2017232)。

摘　　要：目的检测苯乙双胍对胰腺癌及血管生长的抑制作用,分析其抗恶性肿瘤的可能作用机制。方法不同剂量(0.001~10μmol/L)苯乙双胍处理人脐静脉内皮细胞(HUVECs)、人胰腺正常导管上皮细胞系HPDE6-C7和胰腺癌细胞系PANC-1后,CCK-8法检测苯乙双胍对细胞活力的影响;小管形成实验测定苯乙双胍对HUVECs体外血管新生能力的影响;Western blot实验测定血管内皮生长因子A(VEGFA)、p-VEGFR2和p-ERK表达水平。建立原位移植裸鼠胰腺癌模型,检测苯乙双胍对裸鼠胰腺癌生长的作用;明胶-氧化铅血管造影法检测裸鼠体内胰腺肿瘤单位体积血管密度,免疫组化检测CD34和VEGFA的阳性表达。结果0.01~1μmol/L的苯乙双胍可明显抑制PANC-1细胞活力,但对HUVECs和HPDE细胞活力无显著影响;苯乙双胍可抑制HUVECs小管新生,VEGFA可拮抗苯乙双胍对HUVECs小管新生的抑制作用;苯乙双胍可下调PANC-1细胞中VEGFA和p-ERK的表达,可抑制HUVECs中p-VEGFR2的表达。苯乙双胍对裸鼠无明显毒副作用,但可抑制体内胰腺肿瘤的血管新生,并抑制体内胰腺癌增殖及胰腺肿瘤组织中CD34和VEGFA的阳性表达。结论苯乙双胍可明显拮抗体内外胰腺癌生长和血管新生,ERK/VEGFA/VEGFR2可能是其主要抗癌作用机制。Objective To explore the inhibitory effect of phenformin on pancreatic cancer and the angiogenesis,and to analyze the possible mechanism of its anti-tumor effect.Methods After treating human umbilical vein endothelial cells(HUVECs),human pancreatic normal duct epithelial cell line HPDE6-C7,and pancreatic cancer cell line PANC-1 with different doses of phenformin(0.001-10μmol/L),the effect of phenformin on cell viability was detected by CCK-8 method,the effect of phenformin on the angiogenesis of HUVECs in vitro was measured by tubule formation test,and the expression levels of vascular endothelial growth factor A(VEGFA),p-VEGFR2,and p-ERK were measured by Western blot.The orthotopic transplantation model of pancreatic cancer in nude mice was established.The effect of phenformin on the growth of pancreatic cancer in nude mice was detected.The blood vessel density per unit volume of pancreatic tumor in nude mice was measured by gelatin-lead oxide angiography,and the positive expression of CD34 and VEGFA was detected by immunohistochemistry.Results Treatment with 0.01-1μmol/L phenformin significantly inhibited the activity of PANC-1 cells,but had no significant effect on the activity of HUVECs and HPDE cells.Phenformin could inhibit the angiogenesis of HUVECs.VEGFA could antagonize the inhibition of phenformin on the angiogenesis of HUVECs.Phenformin could down-regulate the expression of VEGFA and p-ERK in PANC-1 cells and inhibit the expression of p-VEGFR2 in HUVECs.Phenformin had no obvious toxic and side effects on nude mice,but it could inhibit the angiogenesis of pancreatic tumors in vivo,and inhibit the proliferation of pancreatic cancer in vivo and the positive expression of CD34 and VEGFA in pancreatic tumor tissue.Conclusions Phenformin can significantly inhibit the growth and angiogenesis of pancreatic cancer both in vitro and in vivo,and ERK/VEGFA/VEGFR2 may be its main anti-tumor mechanism.

关 键 词：苯乙双胍 胰腺癌 血管生长 

分 类 号：R735.37[医药卫生—肿瘤]