醋酸丙氨瑞林对人子宫内膜癌裸鼠皮下移植瘤的作用及对TRF2表达的影响

Effect of alarelin acetate on growth of subcutaneous xenograft tumor of human endometrial carcinoma and expression of TRF2 in nude mice

目的探讨不同剂量的醋酸丙氨瑞林对人子宫内膜癌裸鼠皮下移植瘤生长的抑制及端粒重复因子2(telomeric repeat binding factor-2,TRF2)表达的影响。方法选取雌性裸鼠34只,无菌条件下饲养。体外培养人子宫内膜癌HEC-1B细胞株,制成悬液接种于裸鼠皮下组织以建立人子宫内膜癌裸鼠皮下移植瘤模型。选取32只合格的荷瘤裸鼠随机分为4组:对照组(生理盐水肌内注射),低、中、高剂量丙氨瑞林干预组(20μg/kg、40μg/kg、80μg/kg醋酸丙氨瑞林肌内注射),每组8只。观察裸鼠生长发育情况,每7天测量皮下移植瘤体积,4周后处死裸鼠,取出移植瘤,测量肿瘤体积,并绘制生长曲线、计算抑瘤率。移植瘤组织石蜡包埋行HE染色,镜下观察瘤细胞形态变化;免疫组织化学法检测瘤组织中TRF2蛋白的表达情况。结果与对照组比较,丙氨瑞林干预组随药物剂量增加移植瘤体积依次减小(P<0.05);丙氨瑞林低、中、高剂量干预组抑瘤率分别为10.01%、19.80%、44.98%,高剂量组抑瘤率大于低、中剂量组(P<0.05或P<0.01)。HE染色光镜观察见丙氨瑞林干预组瘤细胞排列较对照组稀疏,瘤细胞核深染、固缩。免疫组织化学检测显示,随着药物剂量增加,瘤组织中TRF2蛋白表达逐渐下调(P<0.05)。结论醋酸丙氨瑞林对荷瘤裸鼠子宫内膜癌细胞株HEC-1B皮下移植瘤生长有抑制作用,可能与TRF2蛋白表达下调有关。

Objective To explore the effect of intervention with different doses of alarelin acetate on the growth of subcutaneous xenograft tumor of human endometrial carcinoma and the expression of telomeric repeat binding factor-2（ TRF2） in tumor bearing nude mice. Methods Thirty-four female nude mice were chosen and fed under sterile condition. Human endometrial cancer cell line HEC-1-B cells were cultured in vitro and were inoculated into subcutaneous tissue of nude mice after being made into suspension to establish human endometrial carcinoma xenograft tumor model. Thirty-two qualified tumor bearing nude mice were randomly divided into four groups（ n = 8 each） ： control group（ intramuscular injection of normal saline）,intervention groups with low,medium and high dose of alarelin（ intramuscular injection of 20 μg / kg,40 μg/ kg,80 μg / kg of alarelin acetate）. The growth and development situation of bearing nude mice was observed,and the volume of subcutaneous xenograft tumor was measured once every 7 days. The nude mice were sacrificed 4 weeks late,and the xenograft tumor was taken out,then the tumor volume was measured. The growth curve was drawn,and the tumor inhibition rate was calculated. The xenograft tumor tissues were embedded with paraffin,and HE staining was made to observe the morphological changes of tumor cells by light microscopy. The immunohistochemical method was used to detect the expression of TRF2 protein in tumor tissues. Results Compared with control group,the xenograft tumor volume decreased with the increase of drug doses in alarelin intervention groups（ P〈0. 05）. The tumor inhibition rates in alarelin intervention groups with low,medium and high doses of alarelin were 10. 01%,19. 80%,44. 98%,respectively,and the tumor inhibition rate in alarelin intervention group of high dose was significantly higher than those in alarelin intervention group of low and medium doses（ P〈0. 05 or P〈0. 01）. The HE staining under light microscope observation results showed that compared with control group,the tumor cells presented sparse array,and the cell nucleus presented deep dyeing and shrinkage in alarelin intervention groups. The immunohistochemical method showed that the expression of TRF2 protein was down-regulated gradually with the increase of drug doses（ P〈0. 05）. Conclusion Alarelin acetate has inhibitory effect on growth of subcutaneous xenograft tumor of human endometrial carcinoma cell line HEC-1-B cells in tumor bearing nude mice,and the mechanism may be associated with the down-regulation of TRF2 protein.