黑色素瘤抗原-3基因在肝细胞癌中的研究  

Melanoma antigen 3 expression in human hepatocellular carcinoma

作　　者：蔡胜利[1] 赵海涛[2] 冷希圣[1] 程继华[1] 龚顺友[3] 彭吉润[1] 丛旭[1] 王宇[1] 芮静安[2] 回允中[1] 杜如昱[1] 陈慰峰[3] 

机构地区：[1]北京大学人民医院,100044 [2]邮电总医院肝癌研究所 [3]北京大学基础医学院免疫学系

出　　处：《中华外科杂志》2000年第9期693-696,I037,共5页Chinese Journal of Surgery

基　　金：国家自然科学基金!(39830 42 0 )

摘　　要：目的　检测黑色素瘤抗原 3(melanomaantigen 3 ,MAGE 3)mRNA在肝细胞癌 (HCC)中的表达 ,探讨用MAGE 3基因编码蛋白作为疫苗对HCC患者进行免疫治疗在理论上的可行性。　方法　用RT PCR方法对 45例HCC患者癌组织和相应癌旁肝组织以及 16例肝硬化组织和 12例正常肝组织的MAGE 3mRNA表达情况进行研究 ,对 3例RT PCR扩增产物中目的基因片段进行DNA序列测定 ,并以测序证实为MAGE 3基因的cDNA片段为模板合成 [α32 P]标记的探针 ,进行Southern杂交。同时用ELISA方法对其中 43例HCC患者进行HLA 1 A及B位点分型。　结果　 45例HCC患者中 ,35例 (78% )MAGE 3mRNA表达阳性 ,癌旁组织有 6例阳性 (13% ) ,病理检查证实这 6例癌旁组织中都有明显的异型增生 ,而非HCC患者的肝硬化及正常肝组织均不表达 ;DNA测序 ,3例PCR产物中的目的基因片段均证明为MAGE 3cDNA序列 ;Southern杂交结果和RT PCR电泳结果完全一致 ;HCC患者中常见HLA类型为A2 (5 3 5 % ) ,A11(2 5 6 % ) ,A2 4 (2 0 9% ) ,A33(2 0 9% ) ,B13(2 8 3% ) ,B35(2 3 2 % ) ;MAGE 3的表达与肿瘤直径 ,AFP水平等临床指标无相关关系。　结论　MAGE 3在肝细胞癌中呈高比例表达 ,这使得以此抗原用于HCC患者的免疫治疗成为可能 ;一些细胞有异型增生的癌旁组织中表?Objective[WT5”BZ] To investigate the expression of melanoma antigen 3 (MAGE 3) mRNA in human hepatocellular carcinoma (HCC) and probe into the theoretical feasibility that MAGE 3 antigens can be developed as a new peptide vaccine for immunotherapy in HCC patients. [WT5”HZ]Methods[WT5”BZ] The expression of MAGE 3 mRNA in HCC tissues and the adjacent non HCC liver tissues was studied using RT PCR in 45 HCC patients. The results were compared with those of 16 cirrhotic patients and 12 patients whose liver tissues were pathologically normal. MAGE 3 mRNA positive PCR products were DNA sequenced in 3 HCC patients. The sequenced fragments of MAGE 3 cDNA were used as template by which a [α 32 P] labeled probe was synthesized and employed for Southern blot analysis. HLA class I A and B typing of 43 HCC patients were assayed by ELISA. [WT5”HZ]Results[WT5”BZ] Of the 45 HCC samples, 35 (78%) expressed MAGE 3 mRNA and six HCC adjacent tissues were also positive in MAGE 3 expression. Pathological examination showed cellular heteromorphism in these adjacent tissues.The non HCC liver tissues from chirrhosis and normal liver samples were not MAGE 3 mRNA detectable. The DNA sequence confirmed that the target gene fragment in all of the 3 samples of PCR products was MAGE 3 cDNA. Southern blotting result confirmed that of RT PCR assay. In HCC patients, the predominant types of HLA were A 2 (53 5%), A 11 (25 6%), A 24 (20 9%), A 33 (20 9%), B 13 (28 3%), and B 35 (23 2%). MAGE 3 mRNA expression in HCC showed no correlation with the level of serum AFP and the size of the tumor. [WT5”HZ]Conclusions[WT5”BZ] MAGE 3 mRNA is expressed at a high percentage of HCC samples. This tumor rejection antigen may be used as peptide vaccine for immunotherapy of HCC patients.The phenomena that some non HCC adjacent tissues with heteromorphism can express MAGE 3 like their paired HCC tissues indicate that the expression of MAGE 3 may be an indiacator in the early stage of c

关 键 词：肝细胞癌 黑色素瘤抗原-3 序列分析 DNA 

分 类 号：R735.7[医药卫生—肿瘤]