摘要
目的研究褪黑素(melatonin,MLT)对裸鼠HXO-RB44细胞皮下移植瘤的生长抑制作用及其相关机制。方法50只裸鼠视网膜母细胞瘤(retinoblastoma,RB)皮下移植瘤模型,随机分成5组,每组10只:A组为阴性对照组;B组、C组、D组分别为低、中、高浓度MLT治疗组;E组为阳性对照组。低、中、高浓度MLT治疗组分别按每只每次腹腔注射MLT20 mg.kg-1、40mg.kg-1、60 mg.kg-1各2 mL进行给药,阴性对照组注射等量生理盐水,阳性对照组每只每次腹腔注射等量足叶乙甙(4 mg.kg-1);每天给药1次,共7 d。观察各组裸鼠皮下移植瘤生长情况、裸鼠体质量的变化。采用组织病理学检查,观察裸鼠移植瘤的形态学特征;采用免疫组织化学、Western blotting检测肿瘤细胞血管内皮生长因子(vascular endothelial growth factor,VEGF)、细胞间黏附分子-1(intercellular adhesion molecule-1,ICAM-1)的表达情况。结果E组白细胞、血红蛋白、血小板计数值及谷丙转氨酶、谷草转氨酶、次黄嘌呤核苷值与A组相比,差异均有统计学意义(P均<0.05),MLT各治疗组裸鼠血常规结果及肝、肾功能与A组相比,差异无统计学意义(P>0.05);不同时间点B组、C组、D组裸鼠体质量与A组比较,差异无统计学意义(P>0.05);B组裸鼠皮下移植瘤肿瘤质量与A组相比,差异无统计学意义(P>0.05),表明低剂量的MLT对裸鼠移植瘤的生长无抑制作用,C组、D组与A组进行比较,差异均有显著统计学意义(P均<0.01),表明大剂量的MLT对裸鼠移植瘤的生长有抑制作用;低、中、高浓度MLT治疗组及阳性对照组裸鼠移植瘤抑瘤率分别为12.50%、27.89%、40.72%、44.52%;组织病理学检查发现MLT治疗组及E组肿瘤内出血、坏死的程度明显高于阴性对照组,免疫组织化学及Western blotting检测发现MLT治疗组VEGF、ICAM-1及其蛋白的表达明显低于阴性对照组;MLT的治疗剂量与VEGF、ICAM-1蛋白的表达呈高度负相关。结论MLT对HXO-RB44细胞皮下移植瘤生长有抑制作用;MLT能抑制裸鼠移植瘤内VEGF、ICAM-1蛋白的表达,其浓度与VEGF、ICAM-1蛋白的表达呈高度负相关。
Objective To investigate the inhibitory effects of melatonin（MLT） on HXO-RB44 cells subcutaneous transplantation tumor in nude mice and its related mechanisms.Methods Fifty nude mice models with retinoblastoma（RB） subcutaneous transplantation tumor were randomly divided into 5 groups,10 mice in each group.Group A was negative control group,group B,group C and group D were low,medium and high concentration MLT treatment groups,respectively,and group E was positive control group.Mice of low,medium and high concentration MLT treatment groups were intraperitoneal injected 20 mg·kg-1,40 mg·kg-1 and 60 mg·kg-1 MLT 2 mL,negative control group injected same amount of normal sodium,and positive control group injected same amount of 4 mg·kg-1 etoposide,once daily for 7 days.The growth changes of subcutaneous transplantation tumor and body mass of nude mice were observed;Morphological characteristics of subcutaneous transplantation tumor were observed by histopathological examination;The expression of vascular endothelial growth factor（VEGF） and intercellular adhesion molecule-1（ICAM-1） were detected by immunohistochemistry and Western blotting.Results There were statistical difference for numbers of leucocytes,hemoglobin and thrombocytes and values of glutamic-pyruvic transaminase,glutamic-oxaloacetic transaminase and hypoxanthine in group E compared with those in group A（all P〈0.05）.There was no significant difference for routine blood test results,liver and renal function in MLT treatment groups compared with those in group A（all P〉0.05）;No significant difference was found in body mass of nude mice in group B,group C or group D compared with group A at different times（all P〉0.05）,neither for the weight of subcutaneous transplantation tumor in group B compared with that in group A（P〉0.05）.MLT with low concentration did not inhibit the growth of subcutaneous transplantation tumor in nude mice,and there was significant difference for group C and group D compared with group A（both P〈0.01）.MLT with high concentration inhibit the growth of subcutaneous transplantation tumor.The inhibitory rates for subcutaneous transplantation tumor were 12.50%,27.89%,40.72% and 44.52% in low,medium and high MLT treatment groups and positive control group.Histopathological examination showed that hemorrhage and necrotic degree in MLT treatment groups and positive control group were significantly higher than those in negative control group.The expression of VEGF,ICAM-1 and their proteins in MLT treatment groups were obviously lower than those in negative control group by immunohistochemistry and Western blotting.There was a highly negative correlation of MLT concentration with expression of VEGF and ICAM-1 proteins.Conclusions MLT can inhibit the growth of HXO-RB44 cells subcutaneous transplantation tumor and the expression of VEGF and ICAM-1 proteins in subcutaneous transplantation tumor of nude mice.There was a highly negative correlation of MLT concentration with expression of VEGF and ICAM-1 proteins.
出处
《眼科新进展》
CAS
北大核心
2009年第11期815-820,共6页
Recent Advances in Ophthalmology
