引用本文:肖 啸,任 劲,杨 柒,唐 伟.婴儿双歧杆菌介导HSV-TK/GCV自杀基因治疗系统对裸鼠肾细胞癌肿瘤细胞凋亡及NF-κB活性的影响[J].重庆医科大学学报,2014,38(6):775~780
婴儿双歧杆菌介导HSV-TK/GCV自杀基因治疗系统对裸鼠肾细胞癌肿瘤细胞凋亡及NF-κB活性的影响
Effects of HSV-TK/GCV suicide gene therapy system mediated by bifidobacterium infantis on apoptosis of renal cell carcinoma and activity of nuclear factor kappa B in nude mice
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中文关键词:  婴儿双歧杆菌  肾细胞癌  基因治疗  核因子-κB
英文关键词:bifidobacterium infantis  renal cell carcinoma  gene therapy  nuclear factor kappa B
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肖 啸,任 劲,杨 柒,唐 伟 重庆医科大学附属第一医院泌尿外科重庆 400016 
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中文摘要:
      目的:证实婴儿双歧杆菌(bifidobacterium infantis,BI)介导单纯疱疹病毒胸苷激酶(herpes simplex virus thymidine ki-nase,HSV-TK)/丙氧鸟苷(ganciclovir,GCV)自杀基因系统对裸鼠肾癌的治疗作用,检测其诱导裸鼠肾癌组织细胞凋亡的作用及对核因子-κB(nuclear factor-κB,NF-κB)信号通路的影响。方法:引瘤后原代细胞培养注射细胞悬液法建立裸鼠皮下荷瘤肾癌模型,并随机分为BI-pGEX-TK/GCV组(重组质粒组)、BI-pGEX-5X-1/GCV组(空质粒组)、BI/GCV组(空BI组)和生理盐水/GCV对照组(生理盐水组)。分别经尾静脉注射BI-pGEX-TK、BI-pGEX-5X-1、BI和生理盐水,联合腹腔注射GCV,4周后,称取裸鼠质量及肿瘤质量。TUNEL法检测各组肿瘤细胞凋亡情况,qRT-PCR、Western blot检测各组肿瘤组织Rel A、Caspase-3 mRNA及蛋白表达水平。结果:(1)重组质粒组荷瘤裸鼠体质量明显高于其余3组(P=0.006),肿瘤组织质量明显降低(P=0.003)。(2)TUNEL法检测发现各组均有不同程度的肿瘤细胞凋亡,以重组质粒组最为明显(P=0.000)。(3)与空质粒组、空BI组及生理盐水组比较,重组质粒组Rel A蛋白表达明显下调(P=0.000),Caspase-3蛋白表达明显上调(P=0.000)。结论:BI介导HSV-TK/GCV自杀基因系统对荷瘤裸鼠肾癌有明显的抑制作用,可能通过影响NF-κB信号通路的活性,诱导荷瘤裸鼠肾癌肿瘤细胞的凋亡,发挥抑癌作用。
英文摘要:
      Objective:To confirm effects of bifidobacterium infantis mediated herpes simplex virus thymidine kinase(HSV-TK)/ganci-clovir(GCV) suicide gene system on the treatment of renal cell carcinoma in nude mice and to detect the induction of apoptosis and activaton of nuclear factor-κB(NF-κB) signaling pathway in renal cell carcinoma. Methods:Tumor cells which obtained through di-gestion of subcutaneous tumor were cultured and then subcutaneously injected into the right flank of the test mice.Animals were ran-domly divided into four groups:BI-pGEX-TK/GCV treatment group(recombinant plasmid group),BI-pGEX-5X-1/GCV group (empty plasmid group),BI/GCV group(empty BI group) and normal saline/GCV control group(normal saline group). Either saline or drug was injected through tail vein,followed by intraperitoneal injection of GCV. The quality of life was recorded after 4 weeks of treatment and tumor weight was also evaluated.Apoptosis of tumor was detected by TUNEL assay. Expression of Rel A and Caspase-3 was assessed through qRT-PCR and Western blot. Results:(1)Quality of life of recombinant group was better and tumor weight of recombinant group was significantly lower compared with those of empty plasmid group,empty BI group and normal saline group(P=0.006,P=0.003). (2)Different degrees of apoptosis of tumor cells in each group were detected by TUNEL assay,most obvions in recombinant plasmid group(P=0.000). (3)More importantly,qRT-PCR and Western blot further demonstrated that the recombinant plasmid group had higher level of Caspase-3 protein expression than empty plasmid group,empty BI group and normal saline group(P=0.000),in contrast,the level of Rel A protein expression was the lowest(P=0.000). Conclusion:Bifidobacterium infantis-mediated HSV-TK/GCV suicide gene therapy system induce apoptosis of tumor cells in renal cell carcinoma possibly by affecting the NF-κB signaling pathway,which play a tumor promoter role.
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