蓝舌病毒湖北株对小鼠肾癌细胞体内外的杀伤效应  

Study on the anti-tumor effect of Bluetongue virus-HBC3 on murine renal cell carcinoma cell line Renca in vitro and in vivo

作　　者：杜贤进[1] 张杰[2] 董长垣[2] 周峰[1] 郭应禄[3] 王肖[1] 周晓光[1] 

机构地区：[1]武汉大学人民医院泌尿外科,430060 [2]武汉大学病毒学国家重点实验室 [3]北京大学泌尿外科研究所

出　　处：《中华实验外科杂志》2010年第6期792-794,F0003,共4页Chinese Journal of Experimental Surgery

基　　金：国家高技术研究发展计划（863）资助项目（2006AA02A245）

摘　　要：目的 观察蓝舌病毒湖北株（BTV-HBC3）对小鼠肾癌细胞株Renca体内外的杀伤效应.方法 接种1 MOI的BTV-HBC3于Renca细胞,观察Renca感染后的细胞病变效应（CPE）,应用扫描电镜观察感染48 h后的超微结构,以噻唑蓝（MTT）比色法检测病毒分别以不同感染复数（MOI）感染Renca细胞后24、36、48 h的细胞存活率,并应用琼脂糖凝胶电泳分析感染后48 h的Renca细胞及其对照组病毒基因组.同时制备BALB/C小鼠Renca细胞荷瘤动物模型,观察应用BTV-HBC3处理后皮下荷瘤生长,并进行苏木素-伊红（HE）染色病理分析,应用免疫组织化学检测病毒蛋白的表达.结果 Renca细胞感染BTV-HBC348 h后,CPE程度大于90%,透射电镜下,胞质内可见增殖的病毒颗粒,呈晶格状排列.Renca细胞接种1 MOI的BTV-HBC3 24、36、48 h后细胞的存活率分别为（44.45±5.26）%、（30.47±4.77）%、（20.34±5.26）%,且该病毒对Renca细胞的杀伤作用呈现一定的量效关系;感染后细胞进行琼脂糖电泳可以检测到特异性长（L）、中（M）、短（S）3组分片段的病毒的dsRNA基因组;体内实验表明BTV-HBC3也明显抑制肿瘤生长（P〈0.01）;组织病理学观察中肿瘤组织未见坏死区,可见大片透明空泡样组织,肿瘤组织内可见腺体样结构,而小鼠其他各脏器组织切片中未见病毒感染,治疗组肿瘤组织可见病毒蛋白的阳性染色,其余组织器官经免疫组织化学分析未见病毒蛋白表达.结论 蓝舌病毒湖北株对小鼠肾癌细胞体内外具有显著的靶向杀伤效应.Objective To investigate the anti-tumor effect of Bluetongue virus-HBC3 on murine renal cell carcinoma cell （ Renca） in vitro and in vivo. Methods The renal cells were inoculated with BTVHBC3 at MOI of 1. Observed the cytopathic effects （CPE） by microscopy and the ultra-structural changes by transmission electron. MTT assay was used to comparative analyses of differential survival rates using different Multiplicities of Infection （ MOI） after the renal cells were infected 24,36,48 h. The presence of BTV genome was also detected by agarose gel electrophoresis. The subcutaneous tumor-bearing mice model of renca cells were created up successfully using the BALB/c mice,and the mice was intratumorally injected with BTV-HBC3. Observed the tumor growth, and the pathological and immunohistochemical of the tumor tissue. We also detected the serum IFN-α of the mice. Results Over 90% cytopathic effects （CPE） were readily observed and detected in renca cells infected with BTV 48h postinfection. In electron microscopy （EM）, BTV virions could be seen amplified efficiently in renca cells. The survival rate of the renca cell were respectively （44.45 ± 5. 26） % , （30. 47 ± 4. 77）% , （ 20. 34 ± 5. 26 ） % when infected with BTV at MOI of 1 24,36,48 h postinfection. It showed that there was a dose-effect relationship between them. BTV viral genomic ds-RNA fragments could be found in BTV infected Renca cells through gel electrophoresis. The tumor volume reduced dramatically in contrast with the control group. After BTV treatment Subsequent pathological and immunohistochemical analysis also demonstrated its distinct infection restricted in the Rencacells. Conclusion BTV-HBC3 could selectively degradated Renca cells.

关 键 词：溶瘤病毒 蓝舌病毒 肾细胞癌 

分 类 号：R737[医药卫生—肿瘤]