摘要
背景:胃肠激素蛙皮素作为一种强有力的有丝分裂原,能刺激多种正常和肿瘤组织生长。目的:探讨蛙皮素对人胰腺癌细胞系CFPAC-1的促生长作用及其对细胞周期调控因子细胞周期蛋白(cyelin)D_1和细胞周期蛋白依赖性激酶(CDK)4的调节作用,以及蛙皮素促生长作用的机制。方法:用四唑蓝(MTT)比色法作为增殖指标,研究蛙皮素对CFPAC-1细胞生长的影响。用流式细胞仪观察蛙皮素对CFPAC-1细胞cyclin D_1表达的影响。用Western印迹和增强化学发光法(ECL)自显影观察蛙皮素对CFPAC-1细胞CDK4蛋白表达的影响。结果:一定浓度的蛙皮素对CFPAC-1细胞的生长具有促进作用,以1×10^(-10)mol/L蛙皮素的促生长作用最为明显(MTT OD_(570nm):蛙皮素1×10^(-10)mol/L组和对照组分别为0.652±0.155和0.359±0.109,n=3,P=0.002)。流式细胞仪检测发现,1×10^(-10)mol/L蛙皮素作用30min对CFPAC-1细胞cyclin D_1的表达有促进作用,但无显著统计学差异(P>0.05)。Western印迹和ECL自显影结果表明,1×10^(-10)mol/L蛙皮素作用15min可使CFPAC-1细胞CDK4蛋白表达升高,而后逐渐下降。结论:蛙皮素对CFPAC-1细胞的促生长作用部分是通过蛙皮素介导的细胞周期调控因子cyclin D_1/CDK4蛋白表达升高,从而促进细胞周期进展而实现的。
Background: Bombesin, a kind of gastrointestinal hormone, is a powerful mitogen which can stimulate the growth of normal and cancer cells. Aims: To investigate the growth-stimulating effect of bombesin on human pancreatic cancer cell line CFPAC-1, and the regulatory effect of bombesin on cell cycle regulating factor cyclin D_1 and cyclin-dependent kinase (CDK) 4, and to explore the mechanisms of growth-stimulating effect of bombesin. Methods: The effect of bombesin on the growth of CFPAC-1 cells was studied by using methyl thiazolyl tetrazolium (MTT) as the parameter of cell growth. The effect of bombesin on cyclin D_1 expression of the CFPAC-1 cells was observed by flow cytometry. Western blot and enhanced chemiluminescence (ECL) were performed to identify CDK4 protein expression in CFPAC-1 cells treated by bombesin. Results: Bombesin had significant growth-stimulating effect on CFPAC-1 cells. This effect was more prominent at 1×10^(-10) mol/L in comparison with the controls (MTT OD_(570nm): 0.652±0.155 VS 0.359±0.109, n=3, P=0.002). Flow cytometry demonstrated that bombesin at 1×10^(-10) mol/L increased cyclin D_1 expression in CFPAC-1 cells after 30 minutes exposure compared with the controls (P>0.05). Western blot and ECL showed bombesin at 1×10^(-10) mol/L increased CDK4 protein expression in CFPAC-1 cells after 15 minutes exposure, then the expression decreased gradually. Conclusions: The growth-stimulating effect of bombesin on CFPAC-1 cells is partially mediated by bombesin-mediated cell cycle regulating factor cyclin D_1 and the increased expression of CDK4 protein which in turn promote cell cycle progression.
出处
《胃肠病学》
2003年第3期139-142,共4页
Chinese Journal of Gastroenterology
基金
国家科委九五专项基金
