带有HPV-16 E6/E7基因的DC疫苗抑制裸鼠宫颈癌生长的机制  

The study of inhibitory effect of dendritic cell vaccine carrying HPV-16 E6/E7 gene on the growth of cervical cancer in nude mice and the related mechanisms

作　　者：符少月[1] 焦庆昉[1] 黄启彬[1] 易发平[1] 

机构地区：[1]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医科大学分子医学与肿瘤研究中心,重庆400016

出　　处：《中国老年学杂志》2013年第10期2299-2302,共4页Chinese Journal of Gerontology

基　　金：国家自然科学基金项目(No.30800945)

摘　　要：目的观察由携带HPV-16 E6/E7基因的树突细胞(dendritic cell,DC)疫苗免疫得到的细胞毒性T淋巴细胞(CTL)在裸鼠体内对宫颈癌的抑制作用及相关机制。方法诱导培养C57BL/6小鼠骨髓的未成熟DC(imDC),将已经成功构建的携带人乳头状瘤病毒(HPV)-16 E6/E7基因的重组腺病毒感染imDC,Western印迹检测E7蛋白的表达;构建裸鼠宫颈癌细胞移植瘤模型,应用DC疫苗诱导的CTL处理后,观察裸鼠肿瘤的体积变化;CCK8(cell counting kit8)法检测经DC疫苗免疫的BALB/c小鼠脾脏T淋巴细胞的增殖;免疫组化法检测裸鼠脾脏组织CD4+T淋巴细胞的表达。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-E6E7-DC-CTL-CaSki组与对照组pAd-mock-DC-CTL-CaSki组、DC-CTL-CaSki组、CaSki组相比较,pAd-E6E7-DC-CTL-CaSki组裸鼠肿瘤体积最小(P<0.05);CCK8法检测经DC疫苗免疫的BALB/c小鼠脾脏T淋巴细胞的增殖,结果 pAd-E6/E7-DC组淋巴细胞OD值明显高于pAd-mock-DC组、DC组和PBS组(P<0.01);免疫组化检测各组裸鼠脾脏组织CD4 T淋巴细胞的表达结果显示,pAd-E6E7-DC-CTL-CaSki组CD4表达量最高,pAd-mock-DC-CTL-CaSki组和DC-CTL-CaSki组CD4表达量次之,CaSki组CD4表达量最低。结论带有HPV-16 E6/E7基因修饰的DC疫苗,能够促进小鼠体内T淋巴细胞的增殖,诱导CTL抑制裸鼠宫颈癌移植瘤的生长。Objective To observe the inhibitory action of the spleen cytotoxic T lymphocytes （CTL） obtained from immune mice by dendritic cell vaccine carrying HPV-16 E6/E7 gene on cervical cancer in nude mice and study the related mechanisms. Methods Immature C57BL/6 mice bone marrow- derived DCs （imDCs） were cultured, imDCs were infected with recombinant adenovirus vector carrying human papillomavirus-16 E6/E7 gene. The expression of E7 protein was detected by Western blotting. The nude mice transplantation tumor model of cervical cancer was successfully constructed, and then the models were treated by CTL which were induced by DC vaccine. The tumor vol- ume changes were observed. The proliferation of spleen T lymphocyte of BALB/c mice immunized by DC vaccine was detected by CCK8 （ cell counting kit8 ）. The expression of CD4 ＋ of spleen tissue of nude mice was measured. Results DCs were successfully cultured and DC vaccine was prepared. At 28 d, tumor volume of nude mice was smaller in pAd-E6/E7-DC-T-CaSki group than that in pAd-mock-DC-T-CaS- ki, DC-T-CaSki group and CaSki groups （ P 〈 0.05 ）. The OD values of T lymphocytes of pAd-E6/E7-DC group was significantly higher than that of pAd-mock-DC, DC and PBS groups （P 〈 0. 01 ）. The expression of CD4 ＋ was higher in pAd-E6/E7-DC-T-CaSki group than that in pAd-moek-DC-T-CaSki, DC-T-CaSki and CaSki groups. Conclusions DC vaccine which modified by HPV-16 E6/E7 gene, could promote the proliferation of T lymphocytes in vivo, and inhibit the grow of cervical cancer transplantation tumor in nude mice by induce CTL.

关 键 词：树突细胞 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]