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
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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。Abstract: 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 time- and 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.
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