Chen Guoan, Wang Xiaoju, Yu Jianjun, Varambally Sooryanarayana, Yu Jindan, Thomas Dafydd G, Lin Mu-Yen, Vishnu Prakash, Wang Zhuwen, Wang Rui, Fielhauer Jeff, Ghosh Debashis, Giordano Thomas J, Giacherio Donald, Chang Andrew C, Orringer Mark B, El-Hefnawy Talal, Bigbee William L, Beer David G, Chinnaiyan Arul M
Department of Surgery, University of Michigan Medical School, Ann Arbor, Michigan, USA.
Cancer Res. 2007 Apr 1;67(7):3461-7. doi: 10.1158/0008-5472.CAN-06-4475.
There is considerable evidence that the presence of cancer can elicit a humoral immune response to specific proteins in the host, and these resulting autoantibodies may have potential as noninvasive biomarkers. To characterize the autoantibody repertoire present in the sera of patients with lung adenocarcinoma, we developed a high-density peptide microarray derived from biopanning a lung cancer phage display library. Using a 2,304-element microarray, we interrogated a total of 250 sera from Michigan lung cancer patients and noncancer controls to develop an "autoantibody profile" of lung adenocarcinoma. A set of 22 discriminating peptides derived from a training set of 125 serum samples from lung adenocarcinoma patients and control subjects was found to predict cancer status with 85% sensitivity and 86% specificity in an independent test set of 125 sera. Sequencing of the immunoreactive phage-peptide clones identified candidate humoral immune response targets in lung adenocarcinoma, including ubiquilin 1, a protein that regulates the degradation of several ubiquitin-dependent proteasome substrates. An independent validation set of 122 serum samples from Pittsburgh was examined using two overlapping clones of ubiquilin 1 that showed 0.79 and 0.74 of the area under the receiver operating characteristics curve, respectively. Significantly increased levels of both ubiquilin 1 mRNA and protein, as well as reduced levels of the phosphorylated form of this protein, were detected in lung tumors. Immunofluorescence using anti-ubiquilin 1 antibodies confirmed intracellular expression within tumors cells. These studies indicate that autoantibody profiles, as well as individual candidates, may be useful for the noninvasive detection of lung adenocarcinoma.
有大量证据表明，癌症的存在可引发机体对宿主特异性蛋白质的体液免疫反应，这些产生的自身抗体可能具有作为非侵入性生物标志物的潜力。为了表征肺腺癌患者血清中存在的自身抗体库，我们开发了一种高密度肽微阵列，该微阵列源自对肺癌噬菌体展示文库进行的生物淘选。使用一个包含2304个元件的微阵列，我们检测了来自密歇根州肺癌患者和非癌症对照的总共250份血清，以建立肺腺癌的“自身抗体谱”。在一个由125份血清组成的独立测试集中，发现一组从125份肺腺癌患者和对照受试者血清样本的训练集中得出的22种鉴别肽，能够以85%的灵敏度和86%的特异性预测癌症状态。对免疫反应性噬菌体 - 肽克隆进行测序，确定了肺腺癌中候选的体液免疫反应靶点，包括泛素连接酶1，一种调节几种泛素依赖性蛋白酶体底物降解的蛋白质。使用泛素连接酶1的两个重叠克隆对来自匹兹堡的122份血清样本的独立验证集进行检测，这两个克隆的受试者工作特征曲线下面积分别为0.79和0.74。在肺肿瘤中检测到泛素连接酶1的mRNA和蛋白质水平显著升高，以及该蛋白质磷酸化形式的水平降低。使用抗泛素连接酶1抗体的免疫荧光证实肿瘤细胞内有表达。这些研究表明，自身抗体谱以及单个候选物可能有助于肺腺癌的非侵入性检测。
