内皮素-3对A375人类恶性黑素瘤细胞株骨桥蛋白基因表达的影响  

The Effects of Endothelin-3 on the Expression of Osteopontin Gene in A375 Human Melanoma Cells

作　　者：田进[1] 黄长征[2] 刘业强[2] 李延[2] 陶娟[2] 李家文[2] 涂亚庭[2] 沈关心[3] 

机构地区：[1]华中科技大学同济医学院附属协和医院感染性疾病科,武汉430022 [2]华中科技大学同济医学院附属协和医院皮肤科,武汉430022 [3]华中科技大学同济医学院基础医学院免疫学系,武汉430030

出　　处：《华中科技大学学报（医学版）》2008年第6期720-723,741,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30671891)

摘　　要：目的研究内皮素-3(ET-3)对A375人类恶性黑素瘤细胞株骨桥蛋白(OPN)基因表达的影响。方法体外培养A375细胞株,分别设立对照组,ET-3不同浓度(1、10、100nmol/L)实验组和ET-3同一浓度(100nmol/L)不同时间(6、12、18、24、30、36h)实验组,采用RT-PCR和Western blot结合吸光度分析技术检测骨桥蛋白基因表达情况。结果不同浓度的ET-3能显著上调A375细胞株骨桥蛋白基因表达,与对照组比较差异有统计学意义,并且这种上调效应呈剂量依赖性,不同浓度干预组组间比较差异有统计学意义。在100nmol/LET-3浓度条件下,随着培养时间的延长,OPN的表达逐渐增加,24h达到高峰,随后缓慢下降,RT-PCR结果吸光度分析显示各时间段实验组与对照组比较差异均有统计学意义(均P<0.05),6、12、18、24h实验组组间比较除6h和12h组间差异无统计学意义外,其余各组组间差异均有统计学意义,24、30、36h实验组组间比较差异有统计学意义。Western blot吸光度分析显示各时间段实验组与对照组比较差异有统计学意义,6、12、18、24h实验组组间以及24、30、36h实验组组间比较差异有统计学意义。结论ET-3能时间和剂量依赖性地上调A375细胞株骨桥蛋白基因的表达。Objective Objective To investigate the effects of endothelin-3 （ET-3） on the expression of osteopontin in A375 human melanoma ceils. Methods Cultured cells were incubated with different concentrations of ET-3 for different durations. The RT-PCR and Western blot analyses were used to evaluate the effects of ET-3 on the expression of osteopontin. Results As compared with the control group, different experimental concentrations of ET3 could significantly up-regulate the expression of osteopontin in A375 cell lines （P〈（0.05） in a dose-dependent fashion. There was significant difference in the osteopontin expression among the different concentrations of ET 3 groups （P〈0.05）. Under the concentration of 100 nmol/L ET-3, the expression of osteopontin was increased with the culture time and reached the peak level in 24 h and then decreased gradually. The expression of osteopontin at different time points detected by RT-PCR showed significant difference between ET-3 groups and control group （P〈0.05）. There was significant difference between 6 h, 12 h, 18 h, 24 h ET-3 groups or between 24 b, 30 h, 36 h ET-3 groups （P〈0.05） except the outcomes between 6 h and 12 h groups （P〉0.05）. Similar results were also confirmed by Western blot. Conclusion ET-3 could up regulate the expression of osteopontin gene in A375 human melanoma cells in a dose-and time dependent manner.

关 键 词：黑色素瘤 内皮素3 内皮素受体B 骨桥蛋白 

分 类 号：R349.7[医药卫生—基础医学]