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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 observe the inhibitory effect of anti-insulin-like growth factor-1 receptor (IGF-IR) monoclonal antibody 4F2 combined with cisplatin on the growth of ovarian cancer cells and xenografts. Methods: The expression of IGF-ⅠR in 4 ovarian cancer cells (CAOV3, ES2, SKOV3 and Hey cells) was detected by Western blotting. The binding specificity of anti-IGF-ⅠR monoclonal antibody 4F2 and each ovarian cancer cell line was detected by flow cytometry. The effect of 4F2 on the phosphorylation of IGF-ⅠR in SKOV3 and CAOV3 cells was detected by Western blotting. The inhibitory effect of 4F2 and cisplatin alone or in combination on the proliferation of ovarian cancer cells was detected by CCK-8. To observe the inhibitory effect of 4F2 and cisplatin alone or in combination on the growth of xenografts in nude mice. Results: The expression levels of IGF-ⅠR in SKOV3, CAOV3, ES2 and Hey cells from high to low were not expressed in 4 ovarian cancer cells, 4F2 and SKOV3, CAOV3 and ES2 cells could specifically bind, and 4F2 could inhibit IGF Tyrosine phosphorylation of -I. Cell growth inhibition rate was significantly higher in the combination group with 0.2μg / ml cisplatin than in the 4F2 mAb group [SKOV3: (47.4 ± 3.1)% vs (5.3 ± 0.6)%; t = 3.126, P = 0.007. (46.4 ± 3.1)% vs (30.5 ± 4.1)%; t = 2.933, P = 0.009] and cisplatin group = 0.027. CAOV3: (51.6 ± 2.3)% vs (28.9 ± 2.3)%; t = 3.226, P = 0.034]. In vivo, the inhibitory rate of the combination group on tumor growth in ovarian cancer-bearing mice was also significantly higher than that of the 4F2 mAb [ 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] and cisplatin [SKOV3: (87.3 ± 3.1)% vs (28.9 ± 5.5)%; P = 0.008 7. CAOV3: (86.6 ± 3.5)% vs (32.7 ± 4.1)%; t = 2.91, P = 0.007 3]. CONCLUSION: 4F2 McAb specifically binds to IGF-ⅠR-positive ovarian cancer cells. Combined with cisplatin, it inhibits the growth of ovarian cancer cells and their xenografts.