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.
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
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)
