Screening And Analysis Of Different Expression Gene In Esophageal Squamous Cell Cancer

Objective:Genomics can dynamically be use to detect the completed changes of gene type andquantity in the genesis and progression of neoplasms. Combination of functionalgenomics study and microarray-based gene expression incorporated withbioinformatics is promising to find key genes which control the progression of ESCC,and to make better understand the molecular mechaniasm underlying the ESCCtumorigenesis.Mehods:1, The Whole Human Genome Microarray4x44k v2(Agilent) was carried out on5fresh resected ESCC specimens corresponding with normal mucusal tissue fromESCC patients were obtained for this study. After analying the DEGs by biomedicalsoftware and significant erichment of GO terms between ESCC tissues and normaltissues, the molechular mechnisam underlying the ESCC carcinogensis wasestablished.2, Real time qPCR and IHC were applied to verify the expression of NEK6, acandidate DEG, at mRNA level and at protein level,respectively. After detecting theexpression of NEK6, a gene identified by gene expression microarray, in additional30fresh ESCC samples paired with normal tissue and94cases of formalin-fixed,paraffin embedded (FFPE) ESCC tissues, the correlation between clinicopatholigaical characteristics and the status of NEK6expression was analyzed using medicalstatistics.Results:1, Following the criteria that DEGs at two-fold change with q<0.05or gresterbetween cancerous tissue and normal tissue,1362DEGs including444up-regualtedgenes and918down-regulated genes were identified.2, Of these additional30fresh samples,20(66.7%) showed increased levels ofNEK6mRNA in ESCC tissues (chip fold change=2.60). Meanwhile,49archivedESCC tissue showed high levels of NEK6protein in94cases, and further analysisshowed significant correlation between NEK6protein expression andclinicopatholigial features such as tumor depth (p=0.002),lymph node metastasis(p=0.009), clinical staging (p<0.001) and prognosis (p<0.001) but age, gender orhistological differentiation. The overall5-year survival rate of ESCC patients who hadhigher levels of NEK6protein is dramatically lower than those patients who didn’texpressed. Importantly, NEK6might serve as an independent prognositic factor topredict the prognosis of patients who diagnosed with ESCC.Conclusion:1, Differentitally expressed genes of ESCC tissue can be rapidly detected byhighthrought platform microarray-based gene expression profiling. These genesinvolved in huge changes of ESCC compared with normal tissues at transcriptionallevel, suggesting that further study on the selection of distinct genes would make usbetter understand the molecular mechanism of the carcinogensis of ESCC.2, After screening by gene expression microarray and verifying by real time qPCRand IHC, NEK6, a gene involved in mitosis of cell cycle, showed significantly higherexpression at mRNA level and at protein level. Further study showed that the status ofNEK6protein closely associated with clinicopathological features and prognosis andpatients with negative NEK6expression have a better outcome than the others. Inaddition, NEK6might serve as an independent prognositic factor to predict the prognosis of patients who diagnosed with ESCC. Overview, NEK6, a candidantup-regualted DEG in ESCC tissue, would give us some new clues and thoughts forfurther revealing the pathogenesis of ESCC and view as a potential biomarkers andpromising therapeutic targets for ESCC patients.