非小细胞肺癌p53蛋白及Ser15磷酸化水平检测方法的研究

Detection for p53 protein and Ser15 phosphorylation levels in patients with non-small cell lung cancer

目的:研究人体非小细胞肺癌组织中突变型p53蛋白及p53蛋白15位丝氨酸位点(p53Ser15)的磷酸化状况,p53蛋白、p53Ser15磷酸化与非小细胞肺癌临床病理特征和预后的关系。方法:分别应用酶联免疫吸附实验(ELISA)和免疫组化法检测50例非小细胞肺癌组织、相应正常肺组织及10例肺良性疾病组织中p53蛋白及p53Ser15磷酸化水平。随访观察50例非小细胞肺癌组的转移复发情况。结果:ELISA和免疫组化法均检测到p53蛋白、p53Ser15磷酸化水平在非小细胞肺癌组高于正常肺组织组和肺部良性疾病组,差异有统计学意义(P<0.001,P<0.05)。免疫组化方法检测p53蛋白、p53[pS15]阳性率在淋巴结转移组显著高于无转移组(P<0.05)。随访期内5例出现复发或远处转移的病例p53蛋白、p53[pS15]表达水平高于本组未复发转移病例(P<0.05)。结论:ELISA方法做p53[pS15]、p53蛋白的定量分析是可行的。p53蛋白、p53[pS15]高水平表达可能会有助于肿瘤进展更快。

Objective：To study p53 protein and the phosphorylation status of mutant p53 Ser（serine） 15 loci in non-small-cell lung cancer（NSCLC） tissue samples, and the relationships between p53 protein, the phosphorylation of p53 Ser15 and the clinicopathological characteristics and prognosis of NSCLC patients.Methods：Enzyme Linked-Immuno-Sorbent Assay（ELISA）and Immunohistochemical analysis were used to respectively examined the levels of p53 protein and p53 Ser15 phosphorylation in 50 paired samples obtained from dissected NSCLC and normal neighboring tissue, and 10 benign pulmonary disease samples.At same time metastasis and recrudescence of NSCLC patients was investigated at any time.Results：The levels of p53 protein and p53 [pS15] in NSCLC were significantly higher than those in benign pulmonary disease and normal tissue（p53 P0.001,p53 [pS15]P0.05）.The rates of p53 protein and p53[pS15] positive in lymph nodal involvement was higher than no lymph nodal metastasis （P0.05）.5 NSCLC with metastasis or recurrence exhibited high expression in p53 protein and p53 [pS15] during observation stage （P0.05）.Conclusion：It is possible to quantitative analyse p53 protein and p53 [pS15] by ELISA .High expression in p53 protein and p53 [pS15] may have a growth advantage to tumour cells.