泛素连接酶Cbl-b在黑素瘤组织和细胞系A375、M14、MV3中的表达及意义  

Casitas B-lineage lymphoma b in malignant melanoma tissues and cell lines A375, M14 and MV3：expression and clinical significance

作　　者：王小坡[1] 温斯健[1] 粟倩雅 宋昊[1] 倪娜娜 姜祎群[1] 陈浩[1] 孙建方[1] 

机构地区：[1]中国医学科学院北京协和医学院皮肤病研究所病理科,南京210042 [2]中国医学科学院北京协和医学院皮肤病研究所激光科,南京210042 [3]江苏省皮肤病性病分子生物学重点实验室

出　　处：《中华皮肤科杂志》2016年第12期861-864,共4页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81272992）;江苏省自然科学基金（BK20131063）;北京协和医学院研究生创新基金（2015-1002-01-12）

摘　　要：目的：探讨泛素连接酶Cbl-b在皮肤黑素瘤组织及黑素瘤细胞系A375、M14、MV3中的表达及意义。方法收集69份黑素瘤、30份色素痣组织，采用免疫组化法检测Cbl-b蛋白表达水平。实时荧光定量PCR、免疫印迹法分别检测黑素瘤细胞系A375、M14、MV3及黑素细胞中Cbl-b mRNA及蛋白表达水平。结果69份皮肤黑素瘤标本中，52份（75.36%）表达Cbl-b；30份色素痣标本中，4份（13.33%）表达Cbl-b，两组Cbl-b蛋白表达水平差异有统计学意义（χ2=32.745，P〈0.01）。黑素瘤组织中Cbl-b表达水平与肿瘤进展程度、Clark分级和Breslow厚度均呈正相关（rs分别为0.569、0.654、0.727，均P〈0.01）。实时荧光定量PCR显示，A375、M14、MV3细胞Cbl-b mRNA表达差异有统计学意义（F=176.537，P〈0.01）。免疫印迹法显示，A375细胞Cbl-b蛋白表达水平最高，黑素细胞最低。结论 Cbl-b蛋白在皮肤黑素瘤组织及其细胞株中均高表达。Objective To measure the expression of casitas B-lineage lymphoma b（Cbl-b）in cutaneous malignant melanoma （CMM） tissues and cell lines A375, M14 and MV3, and to assess its clinical significance. Methods Immunohistochemistry was carried out to measure Cbl-b expression in paraffin-embedded tissue sections from 69 cases of CMM and 30 cases of pigmented nevus. Real-time fluorescence-based quantitative PCR and Western-blot analysis were performed to determine the mRNA and protein expressions of Cbl-b in melanocytes and the three cell lines respectively. Results The positive rate of Cbl-b was significantly higher in CMM tissues than in pigmented nevus tissues（75.36%[52/69]vs. 13.33%[4/30], χ2 = 32.745, P 〈 0.01）. The expression level of Cbl-b in CMM tissues was positively correlated with the stage of tumor progression, Clark′s level and Breslow thickness of CMM（rs=0.569, 0.654, 0.727, respectively, all P〈0.01）. As fluorescence-based quantitative PCR showed, there were significant differences in the mRNA expression of Cbl-b among A375, M14 and MV3 cells（F=176.537, P〈0.01）. Western-blot analysis revealed the protein expression of Cbl-b in the 3 melanoma cell lines and melanocytes, which was highest in A375 cells, and lowest in melanocytes. Conclusion Cbl-b was overexpressed in CMM tissues and cell lines.

关 键 词：泛素蛋白连接酶类 黑色素瘤 黑素细胞 肿瘤侵润 肿瘤分级 CBL-B 

分 类 号：R739.5[医药卫生—肿瘤]