肝癌相关抗原血清学自身抗体反应分析

Serological analysis on autoantibody responses to hepatocellular carcinoma assoiated antigens

目的通过9种重组cDNA表达文库血清学鉴定技术(SEREX)筛选的人肝细胞癌(HCC)相关抗原血清学抗体反应进行检测和分析,对是否可作为人肝细胞癌(HCC)诊断标志物进行评价。方法应用重组克隆抗原表达技术表达9种HCC相关肿瘤抗原,免疫印迹法检测HCC、慢性肝炎和正常人血清中自身抗体的反应,并以过9种抗原能否作为诊断HCC的标志物进行分析。结果HCC相关抗原角蛋白23(kerain23)、2a-HS糖蛋白(AHSG)、核糖体蛋白17(RRL17)、铁蛋白轻链(FTL)、RNA解螺旋酶[(DEAD)(Asp-Glu-Ala-Asp box polypeptide 41(DDX41)]在肝细胞患者血清中相应抗体阳性率依次为76.4%,81.8%,78.2%,78.2%,91.0%,慢性肝炎患者血清中相应抗体阳性率依次为60.0%,53.3%,76.7%,56.7%,56.7%,96.7%;正常人血清阳性率为43.3%,46.7%,33.3%,43.3%,76.7%。3种血清抗体阳性率差异有统计学意义的为角蛋白23,a2-HS糖蛋白,核糖蛋白17,铁蛋白轻链,RNA解螺旋酶DDX41;其中肝癌与慢性肝炎和正常人合并阳性率差异有统计学意义的为KRT23,AHSG,ferritin;其他几个相关抗原包括人类骨桥蛋白(Human osteopontin)、丙酮酸脱氢酶激酶同工酶2(pyruvate dehydrogenase kinase,isoenzyme 2)、热休克蛋白P90的三磷酸腺苷酶激活子(activator of heat shock 90kDa protein ATPase)、血浆晒蛋白1(selenoprotein P,plas-ma1)在3种血清中阳性率无差别。结论肝癌相关抗原角蛋白23,a2-HS糖蛋白,铁蛋白轻链有可能成为HCC血清学早期诊断标志物之一。

Objective To investigate the autoantibody responses to 9 HCC-associated antigens which were identified using SERES（serolgical analysis of recombinant cDNA expression library）, and further evaluate whether or not these identified antigens can be used as markers in the detection of HCC. Methods 9 recombinant proteins were expressed from each corresponding cDNA clones which were isolated in previous study, and used as antigens to detect antibodies against each protein in sera from patients with HCC, chronic hepatitis and normal human individuals. Results The frequency of autoantibody against 5 HCC-associated antigens keratin23 （ KRT231, transcript variant 1, a2-HS glycoprotein （AHSG）, ribosomal protein L17 （RRl17 ）, ferritin, light polyteptide （ FTL ）, DEAD （ Asp-Glu-Ala-Asp ） box polypeptide 41 （ DDX41 ） were 46.4 %, 81.8 %,6, 78.2% ,78.2% ,91.0% in HCC60.0%,53.3%,76.7%,56.7%,96.7% in chronic hepatitis, 43.3%, 46.7%, 33.3%, 43.3 %, 76.7% in normal human sera, respectively. There was significant difference in HCC, chronic hepatitis and normal human at KRT23, AHSG, RPL17, FTL, DDX41 ; The significant differcene existed between HCC and merging rate of two the other ones in KRT23, AHSG, FTL. After analyzing the other antigens such as osteopontin, pyruvate dehydrogenase kinase （PDHK）, isoenzyme 2, activator of heat shock 90kDa protein ATPase, selenoprotein P, plasma, 1 （SEPP1）, no significat difference was foud in HCC, chronic hepatitis and normal human. Conclusion The 3 HCC-associated antigens such as KRT23, AHSG, FTL may be one of potential markers in HCC early detection.