干扰素α-1b抑制裸鼠肝异位鼻咽癌种植瘤生长和转移的研究  

Experiment of inhibiting growth and metastasis of xenograft transplanted human nasopharyngeal carcinoma in nude mice by interferon α-1b

作　　者：李湘平[1] 周立辉[1] 刘雄[1] 丁彦青[2] 

机构地区：[1]南方医科大学南方医院耳鼻咽喉科,广州510515 [2]南方医科大学病理教研室

出　　处：《中华耳鼻咽喉头颈外科杂志》2007年第5期372-377,共6页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

基　　金：国家自然科学基金(30471945)

摘　　要：目的观察干扰素α-1b 对裸鼠肝异位鼻咽癌细胞株 CNE-2种植瘤生长和转移的作用并探讨其机理;比较重组腺相关病毒(recombinant adeno-associated virus,rAAV)载体介导干扰素α-1b(interferon-α-1b,IFN-α-1b)基因治疗与 IFN-α-1b 蛋白治疗的优势。方法鼻咽癌细胞株 CNE-2裸鼠肝包膜下注射,建立裸鼠肝脏鼻咽癌异位种植瘤动物模型;采用随机数字表法将40只裸鼠随机分为A～D组,每组10只;24h 后分别经尾静脉注射。A 组:重组腺相关病毒携带人 IFN-α-1b(rAAV-IFN-α-1b),C 组:重组腺相关病毒携带增强型绿色荧光蛋白基因(rAAV-EGFP),D 组:磷酸缓冲液;建模5d 后 B 组:皮下注射入 IFN-α-1b,1次/隔日;3周后每组随机取5只裸鼠断髓处死,观察肝脏成瘤、肺转移及生存期情况,测量肿瘤体积大小,计算抑瘤率,原位末端标记法检测肿瘤细胞凋亡指数;高效液相芯片技术检测外周血中人 IFN-α、小鼠 IL-12的含量,另5只裸鼠观察生存期。结果肝脏鼻咽癌异位种植瘤3周后,肿瘤平均体积(±s)分别为 A 组(0.114±0.116)cm^3、B 组(0.422±0.137)cm^3、C 组(2.476±0.637)cm^3、D 组(2.677±0.704)cm^3,组间比较差异有统计学意义(F=38.536,P<0.01);相对对照 D 组,A 组和 B 组抑瘤率分别为95.74%、84.24%;A、B、C、D 组肺转移率分别为0.0%、0.0%、40.0%、60.0%;凋亡平均指数分别为(21.88±3.29)%、(19.85±1.96)%、(4.37±0.50)%、(3.40±1.05)%,组间比较差异有统计学意义(F=120.964,P<0.01);血清中人 IFN-α平均含量分别为(101.50±11.33)pg/ml、(91.55±9.80)pg/ml、(23.06±4.36)pg/ml、(16.93±9.96)pg/ml,组间比较差异有统计学意义(F=69.128,P<0.01);IL-12平均含量分别为(80.36±13.35)pg/ml、(51.15±9.72)pg/ml、(19.44±7.03)pg/ml、(14.49±4.21)pg/ml,组间比较差异有统计学意义(F=57.116,P<0.01);平均生存期分别为(55.80±2.77)d、(48.20±2.39)d、(35.40±2.61)d、(36.80±1.92)d,组间比较差异有统计学意义(X^2=25.623,P<0.01)。结论 IFN-α-1b 能有�Objective To observe the growth and metastasis effect of interferon α-1b （IFN-α-1b） on nasopharyngeal carcinoma cell line CNE-2 in xenografted model of mice liver. Comparing rAAV-mediated IFN-α-1b gene therapy with the IFN-α-1b protein therapy. Methods The xenografted model of liver nasopharyngeal carcinoma was established by injecting the human nasopharyngeal carcinoma cell line CNE-2 under liver capsule of nude mice. Forty nude mice were randomly divided into four groups by means of random number table method, with ten mice in each one. （Group A： rAAV-IFN-α-1b, Group B： IFN-α-1b; Group C： rAAV-EGFP; Group D： PBS ） . After 24 hours, A, C and D group was injected with rAAV-IFN-α-1b encoding human IFN-α-1b, rAAV-EGFP and phosphate buffer saline via tail vein injection.After 5 days, mice in group B was injected with human IFN-α-1b protein once per two days. Three weeks later five nude mice were sacrificed and then observed their liver tumor formation and pulmonary metastasis. Tumor size was measured and tumor inhibition ratios was calculated, and apoptotic index （AI） was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling （TUNEL） . The contents of human IFN-α contained in peripheral blood and mice IL-12 was determined by high performance liquid chromatography chip techniques. And another five mice were randomly chosen for the observation of surviving study. Results After human nasopharyngeal carcinoma implants in nude mice liver 3 weeks, the average volum of A group（0. 114 ± 0. 116） cm^3 and B group（0. 422± 0. 137） cm^3 were significantly lower than that of C group （2. 476 ±0. 637 ） cm^3 and D group （2. 677± 0. 704） cm^3 （ F = 38. 536, P 〈 0. 01 ）. Compared with D group, the restrained percentage of tumor in group A was 95.74% and group B was 84. 24%. The percentage of lung metastases in group A, B, C and D were 0. 0% ,0. 0% ,40. 0% ,60. 0% respectively. The apoptotic index increased significantly in group A （21.88 ± 3. 29

关 键 词：干扰素Α 依赖病毒 鼻咽肿瘤 

分 类 号：R739.63[医药卫生—肿瘤]