减毒沙门氏菌携带siRNASTAT3质粒对小鼠前列腺移植癌治疗作用.docVIP

减毒沙门氏菌携带siRNASTAT3质粒对小鼠前列腺移植癌治疗作用 　作者：刘喜东 周悦 周姝含 曹群 王芳 魏雁虹 周庆伟 赵雪剑 【摘要】 目的 观察减毒沙门氏菌携带siRNASTAT3质粒对小鼠前列腺移植癌的治疗效果，分析其作用机制。方法 构建小鼠前列腺移植癌模型，随机分组，将转化有siRNASTAT3质粒的减毒沙门氏菌经局部注射到模型构建成功的小鼠体内。观察小鼠一般状态，记录肿瘤体积的变化，RTPCR和Western bolt方法分析STAT3 及其下游基因Bcl2、cMyc、HiF1、cyclinD1的mRNA和蛋白表达水平，铺板法观察菌群分布，HE染色观察肿瘤的形态学变化，免疫组化检查STAT3、HiF1及PCNA蛋白表达情况。结果 ①小鼠前列腺移植癌模型构建成功，携带有siRNASTAT3质粒沙门氏菌治疗组移植性前列腺癌瘤体生长受到明显抑制，瘤重减轻；②铺板法检测菌群分布显示减毒沙门氏菌具有嗜肿瘤特性且在肿瘤内长期存活；肝内分布次之，再次为肺，在脾中分布最低；③pGCSiSTAT3治疗组肿瘤组织的STAT3 mRNA和蛋白表达明显降低；其下游基因Bcl2、cMyc的mRNA及HiF1、cyclinD1蛋白的表达水平都不同程度的受到抑制；④免疫组织化学结果显示pGCSiSTAT3治疗组肿瘤组织的STAT3、HiF1及PCNA表达下调。结论 以减毒沙门氏菌作为运载体携带siRNASTAT3质粒通过调控STAT3下游基因表达，诱导凋亡，对小鼠移植性前列腺癌具有显著的抑瘤作用。 【关键词】 前列腺癌；减毒沙门氏菌；STAT3；RNA干涉 【Abstract】 Objective To observe therapeutic efficacy of attenuated saimonella carrying the siRNAStat3 plasmid towards transplanted prostatic cancer of mouse. Methods The prostatic transplanted model of mouse was built, then the attenuated salmonella with transformed plamid of siRNASTAT3 was injected into local bodies of mouse models constucted successfully. The general status of the mouse were observed and the changing volumes of tumors were recorded.Then the expression of mRNA and protein levels of STAT3 and its downstream genes such as Bcl2,cMyc, HiF1,cyclinD1 were analyzed by RTPCR and Western blot, apoptosis of tumor cells was tested by Flow cytometry.Results (1)Prostatic trnasplanted cancer mondel of mouse were made successfully,and the growth of tumors in the group treated with salmonella of siRNASTAT3 plasmid were suppressed obviously and the weights of tumors were lessened;(2)The apoptotic rates of the cancer group treated with pGCSiSTAT3, pGCSiScramble and group mock were 29.1%,14.7% and 8.5%, respectively. (3)The mRNA and protein expressions of STAT3 in the tumor tissue treated with pGCSiSTAT3 were decreased greatly.The mRNA expressions of downtream genes, such as BcL2,cMyc were inhibited in a different degree, and the protein expressions of HiF1, cyclinD1