Grubbs Elizabeth G, Abdel-Wahab Zeinab, Tyler Douglas S, Pruitt Scott K
Department of General Surgery, Duke University Medical Center, Durham, North Carolina, 27710, USA.
Ann Surg Oncol. 2006 Dec;13(12):1645-54. doi: 10.1245/s10434-006-9029-5.
Real-time quantitative polymerase chain reaction (qPCR) may prove to be a sensitive technique by which to evaluate potential tumor markers in pancreatic cancer.
The prostate stem cell antigen (PSCA) gene was identified as a marker highly expressed in pancreatic adenocarcinoma and not normal pancreas. RNA from pancreatic and nonpancreatic cancer cell lines as well as tissue and blood from pancreatic cancer and control patients was reverse-transcribed and PSCA quantified by qPCR.
Individual operator experience affects the results of qPCR, with significantly different copy numbers at experiment numbers 5, 15, and 40. Five of six pancreatic cell lines had PSCA/actin ratios 10-fold greater than nonpancreatic cancer lines. Mean PSCA expression in pancreatic tumor tissue was significantly higher (P < 0.05, Student's t-test) than in the tissue of benign pancreatic processes. The close correlation of PSCA/actin copy number with number of tumor cells in the blood was demonstrated by regression analysis (r = 0.768, P = 0.0001). PSCA copy number was significantly higher in the blood of patients with metastatic pancreatic cancer than in that of normal patients (P < 0.05, Student's t-test).
Such trends suggest that PSCA may prove to be a valuable pancreatic cancer tumor marker. More generally, the technique of qPCR is shown to provide a sensitive method of evaluating markers in cancer patients.
实时定量聚合酶链反应(qPCR)可能是一种评估胰腺癌潜在肿瘤标志物的敏感技术。
前列腺干细胞抗原(PSCA)基因被鉴定为在胰腺腺癌中高表达而在正常胰腺中不表达的标志物。来自胰腺和非胰腺癌细胞系以及胰腺癌患者和对照患者的组织与血液的RNA进行逆转录,并通过qPCR对PSCA进行定量。
个体操作者的经验会影响qPCR的结果,在实验次数5、15和40时拷贝数有显著差异。六个胰腺癌细胞系中有五个的PSCA/肌动蛋白比率比非胰腺癌细胞系高10倍。胰腺肿瘤组织中的PSCA平均表达显著高于(P<0.05,学生t检验)良性胰腺病变组织中的表达。回归分析表明PSCA/肌动蛋白拷贝数与血液中肿瘤细胞数量密切相关(r=0.768,P=0.0001)。转移性胰腺癌患者血液中的PSCA拷贝数显著高于正常患者(P<0.05,学生t检验)。
这些趋势表明PSCA可能是一种有价值的胰腺癌肿瘤标志物。更普遍地说,qPCR技术被证明是一种评估癌症患者标志物的敏感方法。