成人肝癌组织褪黑素受体表达和结合特性-expression and binding characteristics of melatonin receptor in adult liver cancer tissues.docxVIP

安徽医科大学硕士学位论文 安徽医科大学硕士学位论文 6 6 The difference expression of mRNA in adult hepatoma tissues and its adjacent tissues The mRNA of MR fragment of the expected size in adult hepatoma tissues and its adjacent were determined by RT-PCR. The mRNA of MT1 and MT2 melatonin recep tor subtype were expressed in all groups, but in adult hepatoma had a higher quantity than that in its adjacent tissues(p0.05). The specific binding and kinetic analyses of melatonin receptor in adult hepatoma tissues and its adjacent tissues Specific binding and kinetic analyses of melatonin receptor were measured by radioligand binding assay. By Scatchard analysis, The maximum binding capacity (Bmax) of 125I-Mel specific binding of adult hepatoma tissues and its adjacent tissues was 0.29±0.07pmol · g-1 and 1.16±0.28pmol · g-1(p0.05), Equilibrium dissociation constant (Kd)48.7±6.5 pmol· L-1 and 26.5±4.6pmol· L-1(P0.05). The trait of MR, saturation and reversibility, was detected by 125I-Mel specific binding kinetic analyses. CONCLUSIONS The first found that MR existed in the adult hepatoma tissues, including the two subtypes of MT1 and MT2. The first found that the obvious difference expression of MR and its mRNA in adult hepatoma tissues and its adjacent tissues. MR characters of the specific binding sites were low binding capacity、high affinity、 saturation and reversibility. KEY WORDS adult / hepatoma / melatonin / melatonin receptor / melatonin receptor 1 / melatonin receptor 2 PAGE PAGE 1 英文缩略词表（Abbreviation） 英文缩写 AGE Bmax 英文全名 agarose gel electrophoresis maximum binding capacity 中文全名 琼脂糖凝胶电泳 最大结合容量 β-Actin β-Actin β-肌动蛋白 cAMP cyclic adenosine monophosphate 环磷酸腺苷 cDNA complementary DNA 互补 DNA cGMP cyclic guanosine monophosphate 环磷酸鸟苷 cpm count per minute 每分钟计数 d ay 天 DAB diaminobenzidine 二氨基联苯胺 DEPC diethypyrocarbonate 焦碳酸二乙酯 DMSO dimethyl sulfoxide 二甲基亚砜 EB ethidium bromide 溴化乙锭 FBS fetal bovine serum 胎牛血清 GPCRs G-protein-coupled recepors G 蛋白偶联受体 h hour 小时 HCC hepatocellular carci