Part â… :The Protein Level And Its Significance Of HSP90AB1 In Tumor Tissue And Peripheral Blood Of Lung Cancer Patients Part â…¡:Tumor Driver Gene Target Sequence And Analysis Of LungAdenocarcinoma Tissue

Background and objectiveHSP90AB1 (Heat shock protein 90kDa alpha, class B member 1) is highly conserved ATP-dependent molecular chaperone, and over-expressed in a variety of tumor cells. Some molecules that play important roles in tumor development signaling pathways such as EGFR (epidermal growth factor receptor), HER2 (human epidermal growth factor receptor-2) are HSP90AB1 client proteins. HSP90AB1 interact with these client proteins and participate in a variety of pathophysiological processes of cells. The aim of this study was to detect the expression of HSP90AB1 in non-small cell lung cancer (NSCLC) tissues, and explore its clinical significance.MethodsThe mRNA and protein expression of HSP90AB1 in 62 lung cancer tissues and normal lung tissues was detected by qPCR and Western-blot. The expression of HSP90AB1 in 213 NSCLC tissues and 147 normal lung tissues was detected by tissue microarray and immunohistochemical staining method, and the relationship of HSP90AB1 expression with clinicopathological parameters and prognosis of NSCLC patients were analyzed.The plasma level of HSP90AB1 in 1162 lung cancer patients (include 371 squamous cell carcinoma patients,705 adenocarcinoma patients,80 small cell lung cancer patients) and 282 healthy person was detected by ELISA (enzyme linked immunosorbent assay), the difference level of HSP90AB1 in plasma between lung cancer patients and healthy person were analyzed. And the relationship of HSP90AB1 level in plasma with clinicopathological parameters and prognosis of lung cancer patients were analyzed.ResultsThe mRNA and protein expression level of HSP90AB1 in lung cancer tissues was significantly higher than that in normal lung tissue (P＜0.05). The expression level of HSP90AB1 in lung cancer tissues (positive rate of 54.0%) was significantly higher than that in normal lung tissue (positive rate of 0.0%, P＜0.001). The positive expression rate of HSP90AB1 in lung adenocarcinoma tissues (61.2%) was significantly higher than that in lung squamous cell carcinoma tissues (37.9%) (P=0.002), and its over-expression was associated with poor prognosis in lung adenocarcinoma patients (P=0.032).The expression level of HSP90AB1 had no correlation with clinical stage, lymph node metastasis, pathological grade or other factors (P>0.05).The plasma level of HSP90AB1 in lung cancer patients was significantly higher than that in healthy person (P＜0.001). The plasma level of HSP90AB1 in small cell lung cancer (SCLC) patients was significantly higher than that in non-small cell lung cancer (NSCLC) patients (P＜0.001). The high peripheral blood level of HSP90AB1, CEA (Carcino-embryonic antigen), CA125 (Cancer antigen 125), cyfra211 (Cytokeratin 19 fragments) was associated with poor prognosis in lung cancer patients (P＜0.01), this result was analyzed by Kaplan-Meier method. The high peripheral blood level of HSP90AB1 was associated with poor prognosis in lung cancer patients (P＜0.05), and HSP90AB1 in peripheral blood can be an independent factor to affect the prognosis of patients with lung cancer, this result was analyzed by Cox multi factor analysis method.The plasma level of HSP90AB1 had no correlation with clinical stage, lymph node metastasis, pathological grade or other factors (P>0.05). The plasma of HSP90AB1 can be used as an auxiliary marker in the early diagnosis of lung cancer. The area under the ROC curve (AUC) was 0.888 (95% CI= 0.864-0.909, P ＜0.001), cutoff values was 63.68ng/ml, the sensitivity was 87.8%, specificity was 76.6%.ConclusionHSP90AB1 protein was over-expressed in lung cancer tissues, and was associated with lung cancer pathological type and overall survival in lung adenocarcinoma patients. While the plasma level of HSP90AB1 in lung cancer patients was high, and its high level was correlated with poor prognosis in lung cancer patients. The detection of HSP90AB1, CEA, CA125 and cyfra211 in lung cancer patients’ peripheral blood can be used to determine the prognosis of lung cancer.Background and objectiveLung adenocarcinoma (ADC) is the most common subtype of non-small cell lung cancer. With the development of genetic analysis techniques, the research on the pathogenesis and treatment of tumor is more and more deep. The purpose of this study was to investigate the mutant genes in lung adenocarcinoma tissues, and analyze its clinical significance, based on 483 tumor driver gene sequence of lung adenocarcinoma tumor/normal tissuse.MethodsWe obtained DNA from tumor and matched normal adjacent tissue of forty ADC samples, and carried on quality control. Based on the illumina HiSeq2500 sequencing technology platform build a small fragment library, high coverage of the target area was by captured NovoP products (Agilent sureselectXT Custom 0.5-2.9Mb kit, contains 483 tumor related target gene probe). DNA samples was examined by Paired-End sequencing; sequencing data of 40 cases of tumor samples was obtained. We analyzed the sequencing results of lung adenocarcinoma tumor and matched normal adjacent tissues by using bioinformatics analysis method, and sought for somatic mutations in lung adenocarcinoma tumor and matched normal adjacent tissues.ResultsThere were 31 ADC sample tissuses had reliable exons mutations in the target area, totally 142 (102 genes) mutation positions, and 116 mutation positions (81.69%) were located in the target capture area. Thirtheen high mutation frequency genes were TP53, EGFR, PIK3CA, ATN1, ATXN1, ABL1, ALK, EGF, EPHA3, KRAS, MAP2K3, PIK3CG, and PTPRD. Gene mutations were more frequent in somking and aged patients. We verificated 64 mutant genes by Sanger sequencing, and the validaion rate was 98.4%.ConclusionThese data provided evidence for the further determination of the molecular mechanism and molecular typing of lung adenocarcinoma. Mutation frequency of TP53, EGFR and PIK3CA in lung adenocarcinoma patients was the highest.