促性腺激素释放激素类似物抑制裸鼠卵巢癌移植瘤生长同时保护化疗后卵巢功能的研究

目的 研究促性腺激素释放激素( GnRH)类似物抑制卵巢癌裸鼠皮下移植瘤生长,同时保护化疗裸鼠卵巢功能的双重作用.方法 构建卵巢癌ES-2细胞裸鼠皮下移植瘤模型,随机分为6组(每组6只):(1)生理盐水(NS)组:皮下注射NS 0.1ml/d,1周后腹腔注射NS 0.2ml/周;(2)顺铂(DDP)组:皮下注射NS 0.1ml/d,1周后每周腹腔注射5 mg/kg DDP;(3)戈舍瑞林(goserelin)组:皮下注射goserelin 100 μg/d,1周后腹腔注射NS0.2 ml/周;(4) goserelin+ DDP组:皮下注射goserelin 100 μg/d,1周后每周腹腔注射5 mg/kg DDP;(5)西曲瑞克(cetrorelix)组:皮下注射cetrorelix 100 μg/d,1周后腹腔注射NS0.2 ml/周;(6) cetrorelix+ DDP组:皮下注射cetrorelix 100 μg/d,1周后每周腹腔注射5 mg/kg DDP.用药21 d,观察29 d.比较各组裸鼠体质量、移植瘤体积、移植瘤组织中细胞增殖相关核抗原Ki-67的阳性率、动情周期、卵巢各级卵泡比例、血清抗苗勒管激素(AMH)、卵泡刺激素(FSH)、雌二醇、孕酮水平的差异.结果 各组裸鼠体质量无明显差异(P＞0.05),用药第29天NS组为(19.8 ±2.2)g,DDP组(20.5±1.4)g,gosereline组(19.6±0.9)g,goserelin+DDP组(19.7±1.6)g,cetrorelix组(20.7±2.2)g,cetrorelix+ DDP组(19.0±1.7)g,分别比较,差异均无统计学意义(P＞0.05).用药第12天裸鼠移植瘤体积:NS组为(241±179) mm3,DDP组(78±20) mm3,gosereline组(78±55) mm3,goserelin+ DDP组(64±48) mm3,cetrorelix组(78±64) mm3,cetrorelix+ DDP组(70±19) mm3,用药组均明显小于NS组,差异有统计学意义(P＜0.05);第15、19、22、26、29天各用药组移植瘤体积也均明显小于NS组(P＜0.05).NS组Ki-67阳性率为(33± 10)％,DDP组为3.5％,goserelin组8.8％,goserelin+ DDP组1.5％,cetrorelix组(23±11)％,cetrorelix+ DDP组(8±6)％,DDP组、goserelin组和goserelin +DDP组均明显低于NS组,差异有统计学意义(P＜0.05).goserelin组原始卵泡+窦前卵泡率为(71.5±8.1)％,goserelin+DDP组为(62.4± 4.1)％,cetrorelix组(71.2±7.4)％,cetrorelix+DDP组(63.8±2.9)％,均明显高于DDP组的(47.0±4.8)％,差异均有统计学意义(P＜0.05).goserelin组血清AMH水平为(98±27) ng/ml,明显高于NS组的(66±17) ng/ml,差异有统计学意义(P＜0.05);各组裸鼠血清FSH、雌二醇和孕酮水平分别比较,差异均无统计学意义(P＞0.05).结论 GnRH类似物能抑制卵巢癌裸鼠皮下移植瘤的生长,同时上调AMH分泌、减少动情次数、延长动情周期时间、增加原始卵泡+窦前卵泡率,从而保护卵巢功能.

Study on the effects of gonadotropin-releasing hormone analogues in the inhibition of ovarian cancer transplanted tumors and in the protection of ovarian function after chemotherapy on nude mice

Objective To investigate the influence of gonadotropin-releasing hormone (GnRH) analogues on ovarian cancer and ovarian function in vivo.Methods ES-2 cells were cultured and xenotransplanted into 36 nude mice,which were divided into 6 groups:normal saline (NS) group:NS 0.1 nd/day subcutaneous injection,and then NS 0.2 ml/week peritoneal injection; cisplatin (DDP) group:NS 0.1 ml/day subcutaneous injection,and then DDP 5 mg/kg ( diluted to 0.2 ml ) per week peritoneal injection; goserelin group:100 μg goserelin ( diluted to 0.1 ml) per day subcutaneous injection,and then NS 0.2 ml/week peritoneal injection; goserelin + DDP group:100 μg goserelin ( diluted to 0.1 ml) per day subcutaneous injection,and DDP 5 mg/kg (diluted to 0.2 ml) per week peritoneal injection; cetrorelix group:100 μg cetrorelix (diluted to 0.1 ml) per day subcutaneous injection and NS 0.2 ml/week peritoneal injection; cetrorelix + DDP group:100 μg cetrorelix (diluted to 0.1 ml) per day subcutaneous injection and DDP 5 mg/kg ( diluted to 0.2 ml) per week peritoneal injection.All the peritoneal injection started from subcutaneous injection one week later.To compare the weight of nude mice,the volumes of transplanted tumors,the expression of Ki-67 antigen in transplanted tumors,the estrus,the ratio of atretic follicles,the ratio of primary and preantral follicles,the levels of serum anti-Mullerian hormone ( AMH ),folliclestimulating hormone ( FSH),estradio ( E2 ) and progesterone (P) in each group.Results There were no significant difference in the weight of nude mice among 6 groups ( P ＞ 0.05 ),which on day 29 in NS group was ( 19.8 ±2.2) g,DDP group (20.5 ± 1.4) g,gosereline group ( 19.6 ±0.9) g,goserelin + DDP group ( 19.7 ± 1.6) g,cetrorelix group (20.7 ±2.2) g,and cetrorelix + DDP group ( 19.0 ± 1.7) g.The tumor volumes of different groups on the 12th day:NS group (241 ± 179) mm3,DDP group (78 ±20) mm3,gosereline group (78 t±55) mm3,goserelin + DDP group (64 ±48) mm3,cetrorelix group (78 ±64) mm3,or cetrorelix + DDP group (70 ± 19) mm3,in which there were significant difference between NS group and the other groups ( P ＜ 0.05 ) ; and the same result was obtained on day 15,19,22,26 and 29 ( P ＜ 0.05 ).The expression of Ki-67 in NS group was ( 33 ± 10 ) ％,in which it was higher than those in DDP group 3.5％,goserelin group 8.8％,goserelin + DDP group 1.5％,cetrorelix group (23 ± 11 ) ％,or cetrorelix + DDP group ( 8 ± 6 ) ％ ( P ＜ 0.05 ).The ratio of primary and preantral follicles in goserehn group was (71.5 ± 8.1 ) ％,in goserelin + DDP group was (62.4 ± 4.1 ) ％,in cetrorelix group was (71.2 ± 7.4) ％,and in cetrorelix + DDP group was (63.8 ±3.1 )％,in which they were much higher than that in DDP group ( 47.0 ± 4.8 ) ％ ( P ＜ 0.05 ).The level of AMH in goserelin group was ( 98 ± 27 ) ng/ml,which was much higher than that in NS group (66.2 ± 17.4) ng/ml (P ＜0.05),while there were no difference in the levelsof FSH,E2 or P among different groups ( P ＞ 0.05).Conclusion GnRH analogues could inhibit the growth of transplanted tumors in nude mice,meanwhile increase the secretion of AMH,decrease the frequencies and prolong the lasting time of estrus,decrease the ratio of atretic follicles,raise the ratio of primary and preantral follicles,which may be protect the ovarian function of nude mice.