人乳头状瘤病毒16型多肽疫苗联合紫杉醇＋顺铂化疗方案治疗子宫颈癌的体内外效应研究  

Effect of HPV16 peptide vaccine in combination with paclitaxel-cisplatin chemotherapy on cervical cancer in vitro and in vivo

作　　者：廖书杰[1] 胡晓继[1] 蒋学峰[1] 韩凌斐[1] 夏曦[1] 王薇[1] 王常玉[1] 卢运萍[1] 王世宣[1] 马丁[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院妇产科,武汉430030

出　　处：《中华医学杂志》2010年第43期3035-3039,共5页National Medical Journal of China

基　　金：国家承点基础研究发展“973”计划（2009CB521808）;国家自然科学基金（30901586/C1701）

摘　　要：目的 探讨人乳头状瘤病毒（HPV）16型多肽疫苗联合紫杉醇＋顺铂（TP）化疗方案治疗宫颈癌的可行性,并观察其体内外效应.方法 （1）针对抗原加工相关转运子（TAP）设计HPV16E7蛋白的主要组织相容性复合物Ⅰ（MHC-Ⅰ）抗原结合表位,利用生物信息学分析平台结合体内外实验研究,筛选出一敛性较高、特异性及亲和力较强的HPV16 E7多肽,命名为E7Pa.（2）C57BL/6小鼠注射鼠肺上皮细胞株TC-1细胞（为鼠源性的HPV16阳性的肿瘤细胞株）后,采用等额抽取的随机方法将样本分为6组,E7Pa＋CpG＋TP、E7Pa＋CpG、CpG＋TP、TP和CpG组均为实验组及空白对照组（予生理盐水注射组）.通过定期测量,绘制肿瘤生长曲线,比较各组小鼠肿瘤体积的变化;酶联免疫吸附试验（ELISA）法检测小鼠外周血中γ干扰素（INF-γ）、白细胞介素2（IL-2）的表达水平;TUNEL试剂盒检测肿瘤组织的细胞凋亡情况;记录各组动物自接种肿瘤细胞开始至死亡的时间,绘制生存曲线;作小鼠相关脏器病理检查及白细胞计数观察治疗的安全性.结果 肿瘤生长第60天时,E7Pa＋CpG＋TP组肿瘤体积为（0.013±0.010）cm3与空白对照组（1.900±0.075）cm3相比,差异有统计学意义（P＜0.01）;E7Pa＋CpG组体积为（0.340±0.038）cm3、TP＋CpG组体积为（0.650±0.029）cm3、TP组体积为（1. 100±0.052）cm3,CpG组体积为（0.890±0.047）cm3,分别与空白对照组比较差异有统计学意义（P＜0.05）;小鼠的平均生存时间E7Pa＋CpG＋TP组（108.50±8.97）d,E7Pa＋CpG组（100.02±2.27）d,CpG＋TP组（79.63±4.05）d,TP组（73.24±3.11）d,CpG组（68.63±1.38）d,对照组（52.37±2.47）d,E7Pa＋CpG＋TP组及E7Pa＋CpG组平均生存期与对照组比较生存期明显延长,差异有统计学意义（P＜0.01）;同时,免疫治疗组可有效的激发自身免疫系统活化对抗肿瘤细胞的恶性增生,而化疗组可以显著杀伤恶性细胞,诱导凋亡.Objective To the explore the effect of Human papillomavirus （HPV） 16 peptide vaccine in combination with paclitaxel-cisplatin （TP） chemotherapy on cervical cancer in vitro and in vivo. Methods （ 1 ） The major histocompatibility complex （MHC） class Ⅰ restricted T cell epitopes were studied by bioinformatics for transporter associated with antigen processing （TAP）. Their effects were compared and E7Pa had the most dramatic effect. （2） In vivo, the C57BL/6 mice were divided equally into 6 groups randomly after loading with TC-1 cells （ HPV 16 positive tumor cells from C57BL/6 mouse）,named as E7Pa ＋ CpG ＋ TP, E7Pa ＋ CpG, CpG ＋ TP, TP and CpG group as experiment groups and control （blank injection with physiological saline）. The tumor volumes were measured regularly by tumor growth curve to compare the therapeutic effects in different groups; the related cell factors in murine peripheral blood were evaluated by enzyme-linked immunosorbent assay （ELISA）; the TUNEL test kit was used to explore cellular apoptosis in tumor tissue; the survival curve was drawn from the TC-1 cell loading to natural death; safety was tested by pathological test and leucocyte count. Results At Day 60 of tumor growth, the tumor volume of immunotherapy plus TP chemotherapy group was （0. 013 ±0. 010） cm3 versus the control （ 1. 900 ± 0. 075 ） cm3 with a great significant deviation（ P 〈 0. 01 ）. Meanwhile, the volumes were E7Pa ＋CpG group （0. 340 ±0. 038） cm3 ,TP ＋ CpG group （0. 650 ±0. 029） cm3, TP group （ 1. 100 ±0. 052） cm3 and that of CpG group was （0. 890 ± 0. 047） cm3 separately. And these groups had significant difference with the controls（P 〈0. 05 ）. The average survival time of different groups were E7Pa ＋ CpG ＋ TP group （ 108.50 ±8. 97） d, E7Pa ＋CpG group （ 100. 02 ±2. 27） d, CpG ＋TP group （79. 63 ±4. 05） d, TP group （73. 24 ± 3. 11 ） d, CpG group （ 68. 63 ± 1.38 ） d and controls （ 52.37 ± 2. 47 ） d. And the

关 键 词：人乳头瘤病毒 宫颈肿瘤 疫苗 化疗 

分 类 号：R686[医药卫生—骨科学]