IL-24联合靶向减毒沙门菌载体SL7207/pBud-VP3对胃癌细胞的生长抑制作用  

Inhibiting effect of IL-24 combined with targeting attenuated Salmonella vector SL7207/pBud-Vp3 on growth of gastric cancer cells

作　　者：曹红丹[1] 张朝鸿[1] 吕琳[2] 蔡佩璇[1] 李燕燕[1] 车小雯[1] 

机构地区：[1]重庆医药高等专科学校护理学院,重庆401331 [2]重庆医科大学附属第二医院消化内科,重庆400010

出　　处：《重庆医学》2016年第19期2610-2613,共4页Chongqing medicine

基　　金：重庆医药高等专科学校自然科学启动项目(ygz2013302)

摘　　要：目的探讨白细胞介素(IL)-24联合靶向减毒沙门菌载体SL7207/pBud-VP3对胃癌细胞生长的抑制作用及机制。方法构建pBud-VP3-IL-24真核双表达质粒,利用高压电穿孔法将pBud-Vp3-IL-24质粒导入减毒沙门菌SL7207株中,构成SL7207/pBud-VP3-IL-24菌株。建立小鼠胃癌移植瘤模型,将模型随机分为生理盐水对照组、SL7207/pBud组、SL7207/pBudVP3组及SL7207/pBud-VP3-IL-24组,菌株口服饲喂荷瘤小鼠,测量肿瘤体积并计算抑瘤率。采用蛋白质印迹法检测肿瘤组织中IL-24的表达;采用实时荧光定量-PCR(RT-PCR)检测干扰素-γ(IFN-γ)、IL-6及肿瘤坏死因子-α(TNF-α)水平;采用免疫组织化学法测定半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)及血管内皮生长因子(VEGF)的表达。结果成功构建基于IL-24的减毒沙门菌载体系统SL7207/pBud-VP3-IL-24;治疗14d后,胃癌组织可检测到IL-24蛋白的表达;治疗28d后,SL7207/pBud-VP3-IL-24组与其他组比较,肿瘤体积缩小并能够明显抑制肿瘤生长,差异有统计学意义(P<0.05)。RT-PCR及免疫组织化学显示,IL-24联合SL7207/pBud-VP3能够明显增加免疫因子IFN-γ、IL-6及TNF-α的表达水平(P<0.05),上调Caspase-3及下调VEGF的表达(P<0.05)。结论 IL-24联合SL7207/pBud-VP3可以协同发挥对胃癌细胞的生长抑制作用,其作用机制与肿瘤凋亡促进、肿瘤血管抑制、免疫调节等有关。Objective To investigate the inhibiting effect of interleukin（IL）-24 combined with targeted attenuated Salmonella typhimurium vector SL7207/pBud-VP3 on the growth of gastric cancer ceils. Methods The co-expression eukaryotic expression plasmid pBud-VP3-IL-24 was constructed. The plasmid pBud-VP3-IL-24 was transformed into attenuated Salmonella typhimurium SL7207 by using the high voltage electroporation for constructing the SL7207/pBud-VP3-IL-24 strain. The mouse gastric cancer transplantation tumor model was established and randomly divided into the normal saline control group, SL7207/pBud group, SL7207/pBud-VP3 group and SL7207/pBud-VP3-IL-24 group. The tumor-bearing mice were fed by oral administration of bacterial strain. The tumor volume was measured and the tumor inhibition rate was calculated. The expression of IL-24 was detected by Western blotting. The levels of IFN-7,IL-6 and TNF-~ in tumor tissue were detected by using RT-PCR. The expression of Caspase- 3 and VEGF were detected by using immunohistochemistry. Results The plasmids attenuated Salmonella typhimurium vector carrying the gene IL-24 was successfully constructed. The IL-24 protein expression was detected in gastric cancer tissue after 14 d treatment. The tumor volume after 28 d treatment in the SL7207/pBud-VP3-IL-24 group was reduced compared with the other groups, moreover the tumor growth was significantly inhibited, and the differences were statistically significant（P〈0. 05）. RT-PCR and immunohistochemistry results showed that IL-24 combined with SL7207/bBu＆VP3 could significantly increase the expression levels of immune factor IL-6, IFN-7 and TNF-α in tumor tissue,, in addition,up-regulated the expression of Caspase-3 and down-regulated the VEGF expression（P〈0.05）. Conclusion IL-24 combined with SL7207/pBud-VP3 can synergically play the inhibitory effect on the growth of gastric cancer cells, its mecha- nism is related with the tumor apoptosis promotion,tumor vessel inhibition and immune regulation.

关 键 词：胃癌细胞 白细胞介素-24 减毒沙门菌 

分 类 号：R735.2[医药卫生—肿瘤]