抗IGF-Ⅰ R抗体联合顺铂对卵巢癌细胞及其移植瘤的抑制作用  

Combination anti-tumor effects of an anti-IGF-Ⅰ R antibody and cisplatin on the growth of ovarian carcinoma cells and xenografts

作　　者：张彭南[1] 高慧萍[2] 张鹏飞[2] 孙红[1] 蒋华[2] 

机构地区：[1]复旦大学附属妇产科医院妇科,上海200090 [2]上海交通大学医学院附属仁济医院上海市肿瘤研究所癌基因及相关基因国家重点实验室,上海200032

出　　处：《中国肿瘤生物治疗杂志》2014年第2期136-141,共6页Chinese Journal of Cancer Biotherapy

基　　金：上海市肿瘤研究所硕博创新基金资助(No.SB11-03);高等学校博士学科点专项科研基金资助(No.20100073120092);上海市科委基础研究重点项目资助(NO.12JC1408300)~~

摘　　要：目的:观察抗胰岛素样生长因子Ⅰ型受体(insulin-like growth factor-Ⅰreceptor,IGF-ⅠR)单克隆抗体4F2联合顺铂对卵巢癌细胞及其移植瘤的生长抑制作用。方法:Western blotting检测4种卵巢癌细胞(CAOV3、ES2、SKOV3和Hey细胞)中IGF-ⅠR的表达;流式细胞术检测抗IGF-ⅠR单抗4F2与各卵巢癌细胞系的结合特异性;Western blotting检测4F2对SKOV3和CAOV3细胞内IGF-ⅠR磷酸化水平的影响,CCK-8检测4F2、顺铂单用或联用对卵巢癌细胞的生长抑制作用;建立卵巢癌SKOV3、CAOV3细胞裸鼠移植瘤模型,观察4F2、顺铂单用或联合治疗对裸鼠移植瘤生长的抑制作用。结果:IGF-ⅠR在4种卵巢癌细胞中的表达水平由高到低为SKOV3、CAOV3、ES2,Hey细胞不表达,4F2与SKOV3、CAOV3、ES2细胞均能特异性结合,并且4F2能够抑制IGF-ⅠR的酪氨酸磷酸化。联合组顺铂为0.2μg/ml时细胞生长抑制率即显著高于4F2单抗组[SKOV3:(47.4±3.1)%vs(5.3±0.6)%;t=3.126,P=0.007。CAOV3:(51.6±2.3)%vs(8.2±1.8)%;t=3.516,P=0.009]及顺铂组[SKOV3:(47.4±3.1)%vs(30.5±4.1)%;t=2.933,P=0.027。CAOV3:(51.6±2.3)%vs(28.9±2.3)%;t=3.226,P=0.034];在体内,联合组对卵巢癌种植瘤小鼠肿瘤的抑制率亦显著高于4F2单抗组[SKOV3:(87.3±3.1)%vs(41.6±4.9)%;t=2.29,P=0.004 3。CAOV3:(86.6±3.5)%vs(42.1±7.7)%;t=3.02,P=0.009 1]及顺铂组[SKOV3:(87.3±3.1)%vs(28.9±5.5)%;t=2.56,P=0.008 7。CAOV3:(86.6±3.5)%vs(32.7±4.1)%;t=2.91,P=0.007 3]。结论:4F2单抗特异性结合IGF-ⅠR阳性卵巢癌细胞,联合顺铂能够协同抑制卵巢癌细胞及其移植瘤的生长。Objective： To investigate the antitumor effects of an monoclonal antibody （mAb） against type 1 insulin- like growth factor receptor （ IGF- ⅠR） 4F2 and cisplatin, either each alone or both in combination, on the growth of ovari- an carcinoma cells and xenografts. Methods： Four ovarian carcinoma cell lines （CAOV3, ES2, SKOV3 and Hey cells） were treated with a mAb against human IGF-ⅠR 4F2 and cisplatin, each alone or both in combination. Levels of total IGF-ⅠR protein in CAOV3, ES2, SKOV3 and Hey cells and phosphorylated IGF-ⅠR protein in CAOV3 and SKOV3 cells were analyzed by Western blotting before and after treatments. The binding specificity of the IGF-ⅠR 4F2 antibody to thefour types ovarian carcinoma cells was assessed by flow cytometry. The inhibitory effects of treatments on the growth of o- varian cancer cells was assessed with the CCK-8 assay in vitro. To evaluate these effects in vivo, nude mice were injected with SKOV3 and CAOV3 cells, followed by drug treatment （ i. e. , the IGF-ⅠR 4F2 antibody and 0.2 μg/ml cisplatin, each alone or both in combination）. PBS served as a negative control. The animals were then sacrificed and their ovarian tumor size was examined. Accordingly, rates of the treatment-induced tumor growth inhibition were calculated. Results： IGF- I R protein was detected in CAOV3, ES2 and SKOV3 cells. IGF-ⅠR 4F2 antibody bound specifically to these cells and resulted in a significant decrease in IGF-ⅠR phosphorylation （P 〈 0.05 ）. The combined use of the IGF-ⅠR 4F2 anti- body and cisplatin was significantly more effective than the separate use of these two drugs in inhibiting proliferation in bothSKOV3 cells （[47.4±3.11% vs [5.3 ±0.61% and [30.5±4.11%,P〈0.05） andCAOV3 cells （[51.6± 2. 3 ] % vs [ 8.2 ± 1.8 ] % and, [ 28.9 ± 2.3 ] %, P 〈 0.05）. In vivo, the IGF-ⅠR 4F2 antibody and cisplatin in combi- nation had significantly higher inhibition rates than the two drugs each by itself on tumor growth in both mice injected with SK

关 键 词：胰岛素样生长因子I型受体 4F2 顺铂 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤] R730.51[医药卫生—临床医学]