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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: Gastrointestinal hormone Bombesin, a potent mitogen, stimulates the growth of many normal and tumor tissues. Objective: To investigate the effects of bombesin on the growth of human pancreatic cancer cell line CFPAC-1 and its regulation on the expression of cyelin D_1 and cyclin-dependent kinase (CDK) 4, Mechanism of skin growth promoting effect. Methods: MTT assay was used to evaluate the effect of bombesin on the growth of CFPAC-1 cells. The effect of bombesin on the expression of cyclin D_1 in CFPAC-1 cells was observed by flow cytometry. The effect of bombesin on the expression of CDK4 protein in CFPAC-1 cells was observed by Western blotting and enhanced chemiluminescence (ECL) autoradiography. RESULTS: Bombesin at a certain concentration promoted the growth of CFPAC-1 cells, and the most obvious effect was at 1 × 10 ~ (-10) mol / L of blebbistatin (MTT OD 570nm) 1 × 10 ~ (-10) mol / L group and control group were 0.652 ± 0.155 and 0.359 ± 0.109, respectively, n = 3, P = 0.002). The results of flow cytometry showed that the effect of 1 × 10 ~ (-10) mol / L bombesin for 30 min could promote the expression of cyclin D 1 in CFPAC-1 cells, but there was no significant difference (P> 0.05). The results of Western blot and ECL self-imaging indicated that the expression of CDK4 protein in CFPAC-1 cells was increased after treated with 1 × 10 ~ (-10) mol / L bombesin for 15min, then decreased gradually. CONCLUSION: The effects of bombesin on the growth of CFPAC-1 cells are partially mediated by the increase of the expression of cyclin D 1 / CDK4 protein induced by bombesin and the promotion of cell cycle progression.