CAJ | 학술논문

目的寻找自效的早期诊断及判断肝细胞癌(HCC)预后的血清标志物.方法利用表面增强激光解吸离子化-飞行时间质谱(SELDI-TOF-MS)技术确定HCC患者血清蛋白表达谱.对HCC 81例、肝炎82例和健康志愿者80例血清标本蛋白表达谱进行比较,经高效液相色谱分离和质谱鉴定蛋白质序列.另取48例HCC、54例肝硬化、62例大肠癌、55例鼻咽癌、36例肺癌和42例健康志愿者血清,利用PS20免疫芯片和SELDI技术对候选标志物进行验证.荧光定量PCR技术检测55例HCC患者癌和癌旁组织,13例正常肝组织中候选标志物基因的表达情况,探讨它们与HCC患者临床预后的相关性.结果检出两个HCC候选标志物凝血酶轻链(Thrombin light chain)和生长相关性癌基因编码蛋白(GRO-1).以这两个因子建立的HCC诊断模型敏感性和特异性均为89.6％,显著高于甲胎蛋白(AFP)单独检测敏感性和特异性(69％,83％).它们与AFP联合使用后的灵敏度和特异度更高(91.7％,92.7％),而对大肠癌、鼻咽癌、肺癌的敏感性均不超过50％.HCC 患者癌组织的GRO-1与Thrombin的表达具有显著相关性(r=0.73,P＜0.01),GRO-1表达与肝外转移及术后复发显著相关(P＜0.05),Thrombin的表达量与术后复发及AFP升高相关(P＜0.05).GRO-1和Thrombin高表达的患者术后5年生存率明显低于GRO-l低表达的患者.GRO-1表达量是影响HCC预后的独立影响因素.结论 Thrombin light chain和GRO-1是HCC的潜在血清标志物,并有希望成为HCC患者术后复发转移的预测指标.
목적 심조자효적조기진단급판단간세포암(HCC)예후적혈청표지물.방법 이용표면증강격광해흡리자화-비행시간질보(SELDI-TOF-MS)기술학정HCC환자혈청단백표체보.대HCC 81례、간염82례화건강지원자80례혈청표본단백표체보진행비교,경고효액상색보분리화질보감정단백질서렬.령취48례HCC、54례간경화、62례대장암、55례비인암、36례폐암화42례건강지원자혈청,이용PS20면역심편화SELDI기술대후선표지물진행험증.형광정량PCR기술검측55례HCC환자암화암방조직,13례정상간조직중후선표지물기인적표체정황,탐토타문여HCC환자림상예후적상관성.결과 검출량개HCC후선표지물응혈매경련(Thrombin light chain)화생장상관성암기인편마단백(GRO-1).이저량개인자건립적HCC진단모형민감성화특이성균위89.6％,현저고우갑태단백(AFP)단독검측민감성화특이성(69％,83％).타문여AFP연합사용후적령민도화특이도경고(91.7％,92.7％),이대대장암、비인암、폐암적민감성균불초과50％.HCC 환자암조직적GRO-1여Thrombin적표체구유현저상관성(r=0.73,P＜0.01),GRO-1표체여간외전이급술후복발현저상관(P＜0.05),Thrombin적표체량여술후복발급AFP승고상관(P＜0.05).GRO-1화Thrombin고표체적환자술후5년생존솔명현저우GRO-l저표체적환자.GRO-1표체량시영향HCC예후적독립영향인소.결론 Thrombin light chain화GRO-1시HCC적잠재혈청표지물,병유희망성위HCC환자술후복발전이적예측지표.
Objective To identify potential serum biomarkers specific for hepatocellular carcinoma (HCC).Methods Eighty-one patients wilh hepatitis B-related HCC and 80 healthy controls were randomly divided into a training set (48 HCC,47 controls) and a testing set (33 HCC,33 controls).Serum proteomic profiles were measured using surface-enhanced laser desorption/ionization time-offlight mass spectroscopy (SELDI-TOF-MS).A classification tree was established by the Biomarker Pattern Software.Candidate biomarkcrs were separated by HPLC and identified by MA1DI-MS/MS and database searching.Forty-eight patients with HCC,54 cirrhotic patients and 42 healthy subjects were clinically validated using candidate biomarkers by SELDI-Immunoassay.Real-time reverse transcriptase-polymerase chain reaction was performed to observe GRO-1 and Thrombin in 55 HCC tissues and 13 normal hepatolage tissues.Results Two up-regulated protein peaks were automatically chosen as a classification tree in the training set.These biomarkers were identified as thrombin light chain and CXC chemokines ligand 1 (GRO-1).The sensitivity and specificity of this classification tree were 89.6％.The multivariate model using the two biomarkers and alpha-fetoprotein (AFP) resulted in a sensitivity of 91.7％ and specificity of 92.7％,which was significantly better than AFP alone.The mRNA expression of GRO-1 and Thrombin were found in all HCC tissues.There were significant associations between GRO-1 gene expression and some clinical and pathological findings such as metastasis and recurrence (P＜0.05).Significant differences of 5-year survival rates wee observed among subgroups according to the expression of GRO-1 (P＜0.05).There were significant associations between Thrombin gene expression and some clinical and pathological findings such as recurrence and AFP (P＜0.05).Significant differences of 5-year survival rates were observed among the subgroups according to the expression of THROMBIN (P＜0.05).A positive correlation was found between GRO-1 and Thrombin (r=0.73,P＜0.01).Conclusion Thrombin light chain and GRO-1 are potential biomarkers of HCC.The expression of GRO-1 in HCC tissues was a valuable indicator in estimating metastasis and recurrence in HCC patients.