Humoral immune response to p16, a cyclin-dependent kinase inhibitor in human malignancies.
Humoral immune response to p16, a cyclin-dependent kinase inhibitor in human malignancies.
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DOI：
10.3892/or.16.5.1105
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发表时间：
2006-11
期刊：
影响因子：
4.2
通讯作者：
Koksun Looi;Roxanne Megliorino;F. Shi;Xuan Peng;Yao Chen;Jian-ying Zhang
中科院分区：
文献类型：
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作者：
Koksun Looi;Roxanne Megliorino;F. Shi;Xuan Peng;Yao Chen;Jian-ying Zhang
P16蛋白是一种细胞周期蛋白依赖性激酶(CDK)抑制因子，通过使视网膜母细胞瘤(RB)蛋白磷酸化的细胞周期蛋白依赖性激酶(CDK)失活，在细胞周期调控中发挥重要作用。P16蛋白在多种类型的人类恶性肿瘤中均有过表达。在癌症中对p16的自身抗体反应尚未见报道。这项研究确定了p16自身抗体在不同恶性肿瘤中的范围和频率。在E.Coli BL21(DE3)细胞中表达p16重组蛋白，并用GST融合蛋白纯化系统进行纯化。在进一步的研究中，将p16重组蛋白作为抗原用于酶联免疫分析(ELISA)和Western blotting。对479例癌症患者和82例正常人的血清进行了分析。癌组织中p16自身抗体阳性率为11.7%，与正常人相比差异有统计学意义(p<；0.05)。P16抗体阳性率在鼻咽癌(28.6%)、乳腺癌(17.1%)和肝细胞癌(21.4%)中较高。在56份抗p16抗体阳性的血清中，免疫印迹试验阳性率为85.7%(48/56)。抗原抗体吸附实验也证实了抗p16抗体的特异性。为了提高肿瘤抗体检测的频率，联合使用了p16、p53和c-myc三种肿瘤相关抗原(TAAs)。在乳腺癌、食道癌、鼻咽癌以及肝细胞癌中，p16抗体的出现频率在p<；0.01被发现增加。对于c-myc抗体，在乳腺癌、宫颈癌、结直肠癌和肺癌中发现P<；0.01的频率增加。对于P53抗体，P<；0.01的增加频率仅在乳腺癌中发现。随着三个TAA的相继加入，乳腺癌和鼻咽癌的抗体阳性反应逐步增加，分别达到44%和43%。综上所述，这项研究的结果表明，抗体组合可能获得更高的敏感性，用于早期癌症诊断。可以想象，未来可能会开发出涉及不同TAA面板或阵列的自身抗体图谱，其结果可能对癌症诊断有用。
The p16 protein is a cyclin-dependent kinase (CDK) inhibitor, which plays an important role in the regulation of the cell cycle by inactivating the cyclin-dependent kinase (CDK) that phosphorylates the retinoblastoma (Rb) protein. Overexpression of p16 protein has been found in many types of human malignancy. Autoantibody response to p16 in cancer has not been reported. This study determined the extent and frequency of autoantibodies to p16 in diverse malignancies. p16 recombinant protein was expressed in E. Coli BL21 (DE3) cells, and purified using GST fusion protein purification system. In further studies, p16 recombinant proteins were used as antigens in enzyme-linked immunoassay (ELISA) and Western blotting. Sera from 479 cancer patients and 82 normal individuals were analyzed. Autoantibodies to p16 were found in 11.7% in cancer, with significant difference from the normal individuals (p<0.05). The results in this study also showed that the frequency of antibodies to p16 is relatively higher in nasopharyngeal cancer (28.6%), breast cancer (17.1%) and hepatocellular carcinoma (HCC, 21.4%). Of the 56 ELISA positive sera with the anti-p16 antibodies, 85.7% (48/56) had positive reactions in Western blotting. The antigen-antibody absorption experiment was also performed to confirm the specificity of the anti-p16 antibody. In order to increase the frequency of antibody detection in cancer, a combination of three tumor-associated antigens (TAAs) p16, p53 and c-myc were used. Increased frequencies at p<0.01 were found for antibodies to p16 in breast, esophageal, and nasopharyngeal cancer as well as HCC. For antibodies to c-myc, increased frequencies at p<0.01 were found in breast, cervical, colorectal and lung cancer. For antibodies to p53, increased frequencies at p<0.01 were only found in breast cancer. With the successive addition of three TAAs, there was a stepwise increase of positive anti-body reaction up to 44% in breast cancer and 43% in nasopharyngeal cancer. In summary, the results in this study suggest that the combination of antibodies might acquire higher sensitivity for early cancer diagnosis. It is conceivable that auto-antibody profiles involving different panels or arrays of TAAs might be developed in the future and the results could be useful for cancer diagnosis.