铜绿假单胞菌注射液诱导人胰腺癌细胞PANC-1的凋亡  

Experimental Study on Inducing Apoptosis of Human Pancreatic Carcinoma Cells PANC-1 by Pseudomonas Aeruginosa Vaccinein in vitro

作　　者：包丽颖[1] 孙建民[1] 张玉宝[1] 

机构地区：[1]哈尔滨医科大学附属第三医院肝胆胰外科,哈尔滨150000

出　　处：《肿瘤防治研究》2013年第7期643-647,共5页Cancer Research on Prevention and Treatment

摘　　要：目的观察铜绿假单胞菌注射液对体外培养的PANC-1细胞生长抑制及凋亡的影响。方法体外培养人胰腺癌细胞PANC-1,采用MTT法检测不同浓度PA-MSHA(铜绿假单胞菌)对PANC-1细胞的抑制情况,透射电子显微镜观察药物作用后细胞超微结构变化,TUNEL法观察细胞凋亡,采用流式细胞术检测PA-MSHA与对照组(药物浓度为0)的细胞凋亡率,Westen blot检测促凋亡蛋白caspase-3,caspase-7,caspase-8,caspase-9,PARP的表达。结果 MTT观察到不同浓度PA-MSHA(10×108/ml,5×108/ml,2.5×108/ml,1.25×108/ml,0.625×108/ml)分别作用24、48、72 h后和对照组相比(P=0.006)可使PANC-1细胞生长明显抑制,抑制率呈浓度、时间依赖关系;电子显微镜下PA-MSHA作用后形成凋亡小体,细胞内空泡形成;TUNEL法观察细胞核深染,细胞发生早期凋亡。流式细胞术观察PA-MSHA处理组(2.5×108/ml)与对照组分别作用48 h后处理组的细胞凋亡率明显增高,PA-MSHA作用24 h后和对照组相比,随药物浓度增高,凋亡相关蛋白caspase-3,caspase-7,caspase-8,caspase-9,PARP表达随之增强。结论 PA-MSHA可诱导人胰腺癌细胞PANC-1细胞凋亡进而抑制肿瘤生长。Objective To investigate the inhibitory effects of Pseudomonas aeruginosa vaccine （PA- MSHA） on the proliferation of human pancreatic cancer cells and explore the possible mechanism. Methods MTT assay was used to determine the cellgrowth of human pancreatic cancer cell line panc-1 in vitro treated with the vaccine. Inhibition of cellproliferation,changes of super-microstructure and apoptosis of panc-1 were observed by scanning electron microscopy and Tunel technique. The apoptosis rates of the cells from the PA-MSHA group and the control group （the drug concentration was 0） were detected by flow cytometry. The expression levels of apoptosis proteins were evaluated by Western blot. Results PA-MSHA treatment significantly suppressed the proliferation of panc-1 cells from the PA-MSHA group and the control group in a timeand concentration-dependent manner compared with the control group （P = 0. 006）. PA-MSHA vaccine could induce the decrease in number and size, destruction of cell membrane an microvilli, vacuole in cytoplasm and deep staining of nucleus. The expressions of the apoptosis protein caspase-3, caspase-7, caspase-8, caspase-9 and PARP up-regulated in PA-MSHA-treated cells. Conclusion PA-MSHA can suppress the proliferation of pancreatic carcinoma cell in vitro by promoting cell apoptosis.

关 键 词：胰腺癌 凋亡 PANC-1 铜绿假单胞菌 

分 类 号：R735.9[医药卫生—肿瘤]