IPDMS Based Microarray Technology Apply To The Clinical Research Of Non-small Cell Lung Cancer

Objective To investigate the application of iPDMS based peptide microarray and protein microarray in the clinical research of non-small cell lung cancer.Methods First part p53 and extracellular domain of EGFR were cut into overlapped peptides to build a high-content precise peptide library and then measuring scheme of i PDMS based peptide microarraywasestablished;p53 and EGFR autoantibodies of 150 NSCLC patients and 96 healthy controls were detected by peptide microarray; Traditional bioinformatics tools were used to characterize the linear antigenic regions of p53 and EGFR for further analysis.Second part The levels of serum IL-6、IL-8、MCP-1and GM-CSF of 50 NSCLC patients and 50 healthy controls were measured by i PDMS based protein microarray. Clinical pathological parameters were analyzed at the same time.Results First part Autoantibodies to p53 and EGFR peptides were present in serum of NSCLC patients and healthy controls.4 p53 peptides and 2 EGFR peptides with significant different response in NSCLC and healthy samples were extracted by significance analysis of microarrays(SAM), autoantibody response to these 6 peptides in NSCLC group were significantly higher than healthy group(P<0.05).Receiver operator characteristic curve(ROC) analysis suggests that a combined diagnosis model of p53-02, p53-04, p53-05 and EGFR-48 is most efficient at discriminating NSCLC from healthy controls, which was found to have an overall sensitivity of 36.7% at specificity of 90.0%.Several linear antigenic sites were proved to be within the amino and carboxyl terminal of p53 and the L2 and S2 domain of EGFRSecond part Compared with healthy controls, serum IL-6、IL-8、and GM-CSF levels increased evidently(P<0.01), while serum MCP-1 levels increased(P<0.05) in NSCLC patients. Compared with stage I+II NSCLC patients, serum IL-6 levels of stage III+IV NSCLC patients increased evidently(P<0.01), serum MCP-1 levels of stage III+IV NSCLC patients decreased(P<0.05),serum GM-CSF levels of stage III+IV NSCLC patients decreased evidently(P<0.01),there were no significant difference in serum IL-8 levels(P>0.05). The levels of serum IL-6 of NSCLC patients with lymph node metastasis was significantly higher than those without(P<0.05),and there were no significant difference in serum IL-8、MCP-1 and GM-CSF levels(P>0.05). The levels of serum IL-8 of adenocarcinoma patients was significantly higher than those of squamous carcinoma patients(P<0.05),and there were no significant difference in serum IL-6、MCP-1 and GM-CSF levels(P>0.05).Conclusions1. Autoantibodies to p53 and EGFR peptides differentially expressed between NSCLC group and healthy group were identified by i PDMS based peptide microarray. The combined diagnosis model consists of p53 and EGFR peptides shows promising future for further application. i PDMS based peptide microarray profiling provides a powerful tool to study the precise linear epitope of p53 and EGFR.2. The changes of serum IL-6、IL-8、MCP-1 and GM-CSF levels in NSCLC patients may be closely related to the progression of NSCLC, their serological quantitatively tests helps in investigating immune function of NSCLC patients and could be a useful reference in distinguishing disease stage and tissue type.3. i PDMS based microarray technology,with the advantage of miniaturization、high throughput、multiple indicator and lower cost, provides convenience for immunological research and shows promising future for scientific research and clinical implication.