中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2010年
43期
3035-3039
,共5页
廖书杰%胡晓继%蒋学峰%韩凌斐%夏曦%王薇%王常玉%卢运萍%王世宣%马丁
廖書傑%鬍曉繼%蔣學峰%韓凌斐%夏晞%王薇%王常玉%盧運萍%王世宣%馬丁
료서걸%호효계%장학봉%한릉비%하희%왕미%왕상옥%로운평%왕세선%마정
人乳头瘤病毒%宫颈肿瘤%疫苗%化疗
人乳頭瘤病毒%宮頸腫瘤%疫苗%化療
인유두류병독%궁경종류%역묘%화료
Human papillomavirus%Cervix neoplasm%Vaccine%Chemotherapy
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.免疫治疗与化疗的联用明显优于两者单用,可以在充分调动自身免疫杀伤效应的同时增加化疗药物的抗肿瘤效应.在药物副作用的研究中,联合用药与其他实验组差异无统计学意义.结论 在动物模型中,人乳头状瘤病毒16型多肽疫苗联合紫杉醇+顺铂化疗可以有效治疗HPV16 E7阳性的肿瘤.
目的 探討人乳頭狀瘤病毒(HPV)16型多肽疫苗聯閤紫杉醇+順鉑(TP)化療方案治療宮頸癌的可行性,併觀察其體內外效應.方法 (1)針對抗原加工相關轉運子(TAP)設計HPV16E7蛋白的主要組織相容性複閤物Ⅰ(MHC-Ⅰ)抗原結閤錶位,利用生物信息學分析平檯結閤體內外實驗研究,篩選齣一斂性較高、特異性及親和力較彊的HPV16 E7多肽,命名為E7Pa.(2)C57BL/6小鼠註射鼠肺上皮細胞株TC-1細胞(為鼠源性的HPV16暘性的腫瘤細胞株)後,採用等額抽取的隨機方法將樣本分為6組,E7Pa+CpG+TP、E7Pa+CpG、CpG+TP、TP和CpG組均為實驗組及空白對照組(予生理鹽水註射組).通過定期測量,繪製腫瘤生長麯線,比較各組小鼠腫瘤體積的變化;酶聯免疫吸附試驗(ELISA)法檢測小鼠外週血中γ榦擾素(INF-γ)、白細胞介素2(IL-2)的錶達水平;TUNEL試劑盒檢測腫瘤組織的細胞凋亡情況;記錄各組動物自接種腫瘤細胞開始至死亡的時間,繪製生存麯線;作小鼠相關髒器病理檢查及白細胞計數觀察治療的安全性.結果 腫瘤生長第60天時,E7Pa+CpG+TP組腫瘤體積為(0.013±0.010)cm3與空白對照組(1.900±0.075)cm3相比,差異有統計學意義(P＜0.01);E7Pa+CpG組體積為(0.340±0.038)cm3、TP+CpG組體積為(0.650±0.029)cm3、TP組體積為(1. 100±0.052)cm3,CpG組體積為(0.890±0.047)cm3,分彆與空白對照組比較差異有統計學意義(P＜0.05);小鼠的平均生存時間E7Pa+CpG+TP組(108.50±8.97)d,E7Pa+CpG組(100.02±2.27)d,CpG+TP組(79.63±4.05)d,TP組(73.24±3.11)d,CpG組(68.63±1.38)d,對照組(52.37±2.47)d,E7Pa+CpG+TP組及E7Pa+CpG組平均生存期與對照組比較生存期明顯延長,差異有統計學意義(P＜0.01);同時,免疫治療組可有效的激髮自身免疫繫統活化對抗腫瘤細胞的噁性增生,而化療組可以顯著殺傷噁性細胞,誘導凋亡.免疫治療與化療的聯用明顯優于兩者單用,可以在充分調動自身免疫殺傷效應的同時增加化療藥物的抗腫瘤效應.在藥物副作用的研究中,聯閤用藥與其他實驗組差異無統計學意義.結論 在動物模型中,人乳頭狀瘤病毒16型多肽疫苗聯閤紫杉醇+順鉑化療可以有效治療HPV16 E7暘性的腫瘤.
목적 탐토인유두상류병독(HPV)16형다태역묘연합자삼순+순박(TP)화료방안치료궁경암적가행성,병관찰기체내외효응.방법 (1)침대항원가공상관전운자(TAP)설계HPV16E7단백적주요조직상용성복합물Ⅰ(MHC-Ⅰ)항원결합표위,이용생물신식학분석평태결합체내외실험연구,사선출일렴성교고、특이성급친화력교강적HPV16 E7다태,명명위E7Pa.(2)C57BL/6소서주사서폐상피세포주TC-1세포(위서원성적HPV16양성적종류세포주)후,채용등액추취적수궤방법장양본분위6조,E7Pa+CpG+TP、E7Pa+CpG、CpG+TP、TP화CpG조균위실험조급공백대조조(여생리염수주사조).통과정기측량,회제종류생장곡선,비교각조소서종류체적적변화;매련면역흡부시험(ELISA)법검측소서외주혈중γ간우소(INF-γ)、백세포개소2(IL-2)적표체수평;TUNEL시제합검측종류조직적세포조망정황;기록각조동물자접충종류세포개시지사망적시간,회제생존곡선;작소서상관장기병리검사급백세포계수관찰치료적안전성.결과 종류생장제60천시,E7Pa+CpG+TP조종류체적위(0.013±0.010)cm3여공백대조조(1.900±0.075)cm3상비,차이유통계학의의(P＜0.01);E7Pa+CpG조체적위(0.340±0.038)cm3、TP+CpG조체적위(0.650±0.029)cm3、TP조체적위(1. 100±0.052)cm3,CpG조체적위(0.890±0.047)cm3,분별여공백대조조비교차이유통계학의의(P＜0.05);소서적평균생존시간E7Pa+CpG+TP조(108.50±8.97)d,E7Pa+CpG조(100.02±2.27)d,CpG+TP조(79.63±4.05)d,TP조(73.24±3.11)d,CpG조(68.63±1.38)d,대조조(52.37±2.47)d,E7Pa+CpG+TP조급E7Pa+CpG조평균생존기여대조조비교생존기명현연장,차이유통계학의의(P＜0.01);동시,면역치료조가유효적격발자신면역계통활화대항종류세포적악성증생,이화료조가이현저살상악성세포,유도조망.면역치료여화료적련용명현우우량자단용,가이재충분조동자신면역살상효응적동시증가화료약물적항종류효응.재약물부작용적연구중,연합용약여기타실험조차이무통계학의의.결론 재동물모형중,인유두상류병독16형다태역묘연합자삼순+순박화료가이유효치료HPV16 E7양성적종류.
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 difference between the E7Pa + CpG + TP and E7Pa + CpG groups had great significance with the controls ( P ＜ 0. 01 ).Furthermore, the immune system was effectively stimulated for suppressing tumor growth in the immunotherapy group while cell apoptosis was significantly induced in the chemotherapy group. The combination of immunotherapy and chemotherapy was significantly efficacious than either of them alone. And it could thoroughly stimulate the immune effects and enhance the anti-tumor function of chemotherapeutical drugs. In safety test, there was no significant difference among all groups. Conclusion The HPV16 peptide vaccine in combination with TP chemotherapy can treat the HPV16 E7 positive tumor effectively in experiment.
