带有HPV-16 E6/E7基因的树突状细胞疫苗对裸鼠人宫颈癌CaSki细胞移植瘤的抗肿瘤作用  

Anti-tumor effects of dendritic cell vaccine carrying HPV-16 E6/E7 gene on the transplantable tumor of human cervical cancer in nude mice

作　　者：符少月[1] 焦庆昉[1] 黄启彬[1] 刘潇淇[1] 易发平[1] 宋方洲[1] 

机构地区：[1]重庆医科大学分子医学与肿瘤研究中心重庆医科大学基础医学院生物化学与分子生物学教研室,重庆400016

出　　处：《重庆医科大学学报》2013年第5期512-516,共5页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:30800945);重庆市科委自然科学基金资助项目(编号:CSTC2008BB5225)

摘　　要：目的:探索带有HPV-16 E6/E7基因的树突状细胞(dendritic cell,DC)疫苗对裸鼠人宫颈癌CaSki细胞移植瘤的抗肿瘤作用。方法:体外诱导培养C57BL/6小鼠骨髓的未成熟DC(immature DC,imDC),将已经成功构建的携带人乳头状瘤病毒(human papillomavirus,HPV)-16 E6/E7基因的重组腺病毒感染imDC,Western blot检测E7蛋白的表达;构建裸鼠的人宫颈癌CaSki细胞移植瘤模型,应用DC疫苗诱导的BALB/c小鼠脾脏的细胞毒性T淋巴细胞(cytotoxic T lymphocytes,CTL)处理后,观察裸鼠肿瘤体积变化,肿瘤长出第28天后处死裸鼠,小心剥下肿瘤组织,石蜡包埋,HE染色观察各组肿瘤的细胞形态,免疫组织化学技术法检测肿瘤组织中Bcl-2、Bax蛋白的表达情况。结果:成功培养出DC并制备出DC疫苗;第28天各组裸鼠肿瘤体积pAd-E6/E7-DC-CTL-CaSki组与pAd-mock-DC-CTL-CaSki组、DC-CTL-CaSki组、CaSki组相比较,pAd-E6/E7-DC-CTL-CaSki组裸鼠肿瘤体积最小(P<0.05);免疫组化检测肿瘤组织中Bax蛋白的表达结果显示pAd-E6/E7-DC-CTL-CaSki组最高,而CaSki组表达最低;免疫组化检测肿瘤组织中Bcl-2蛋白的表达结果显示pAd-E6/E7-DC-CTL-CaSki组最低,而CaSki组表达最高。结论:带有HPV-16 E6/E7DC疫苗能够诱导CTL抑制裸鼠人宫颈癌CaSki细胞移植瘤的生长。Objective ：To explore the anti-tumor effects of dendritic eell（DC） vaccine carrying HPV-16 E6/E7 gene on the CaSki cell transplantable tumor of human cervical cancer in nude mice. Methods ：Immature C57BL/6 DC（imDC） was cultured and was infected with recombinant adenovirus vector carrying human papillomavirus-16 E6/E7 gene. Expressions of E7 protein were detected by West- ern blot. CaSki cell transplantable tumor model of human cervical cancer was constructed in nude mice and was treated by cytotoxic T lymphocytes（CTL） ,acquired from BALB/c mice. Changes in tumor volume were observed. Nude mice were sacrificed and Paraffin- embedded on the 28th d since tumors appeared. Tumor cell morphology was observed in each group by ttE staining. Expressions of Bcl-2 and Bax protein of tumor tissues were detected by immunohistochemical techniques. Results ： DC was successfully cultured and DC vaccine was prepared. On the 28th d,tumor volume of nude mice was smaller in pAd-E6/E7-DC-CTL-CaSki group than in pad- mock-DC-CTL-CaSki group, DC-CTL-CaSki group and CaSki group（P〈O.05）. Bax protein expressions of tumor tissues detected by immunohistochemical techniques demonstrated that：expressions in pAd-E6/E7-DC-CTL-CaSki group were the highest while in CaS- ki group were the lowest. Bcl-2 protein expressions of tumor tissue detected by immunohistochemical techniques demonstrated that： expressions in pAd-E6/E7-DC-CTL-CaSki group were the lowest while those in CaSki group were the highest. Conclusions：DC vac- cine modified by HPV-16 E6/E7 gene can inhibit the grow of cervical cancer transplantable tumor in nude mice by inducing CTL.

关 键 词：树突状细胞 人乳头瘤病毒16 T淋巴细胞 CASKI细胞 

分 类 号：R737.33[医药卫生—肿瘤]