Screening And Analysis Of Highly Associated Genes Of Early Esophageal Squamous Cell Carcinomas In Kazak In Xinjiang

Objective:The living environment and lifestyle of Kazak in Xinjiang have been changed,but the incidence and mortality of esophageal carcinoma remains at high level.The key reason is hard to early detect and its prognosis is also poor,the 5-year survival for EC patients low as 10%.At present early diagnosis of EC is by means of endoscopic biopsy.Biopsy depends on pathology as a qualitative detection,but it is failue to accurately diagnosis precancerous lesion due to the limitation of the pathological method.Recognizing precancerous lesions progress to cancer is the vital event.The current researches highlighted that multiple genetic alterations,especially for loss of tumor suppressor genes and activation of oncogenes,were the key factor leading to dysplasia and then carcinogenesis of EC.Therefore,by analyzing differential expression of genes in esophageal precancerous lesions,the highly associated genes for early genetic diagnosis could be determined,which will improve endoscopic diagnosis specificity and remedy the defect of pathology.The morphology and molecular biology of EC had been extensively studied,and now it is generally accepted that the changes of esophageal adjacent cancer tissues stand for early esophageal carcinoma.In this study RT-PCR,Western blot,immunohistochemistry and topology were used to analyze differential expression genes between adjacent carcinoma tissues and distant non carcinoma tissues in esophageal squamous cell carcinoma of Kazak in Xinjiang.We tried to establish highly associated genes in carcinogenesis and progression of ESCC.Moreover,two topology maps were constructed by different group genes,one was from this study,the other was obtained from literature which twenty proteins were reported to demonstrate abnormal expression levels in the early stage of ESCC in other national region.This study will provide the laboratory basis and theoretical evidence on early genetic diagnosis ESCC of Kazak in Xinjiang.Futhermore,applying them as early genetic diagnosis biomarkers in clinic will greatly enhance the diagnosis specificity of biopsy.Meanwhile it will further lay a foundation for genetic pathogenesis of ESCC.Methods:(1)Semi-quantitative RT-PCR was used to detect 42 candidate genes mRNA levels of ESCC tissues and distant non carcinoma tissues among 48 cases of ESCC in Kazak,and screened for the highly related genes by the ratio in mRNA levels.Highly related genes further detected in adjacent carcinoma tissues and distant non carcinoma tissues among 10 cases of ESCC by semi-quantitative RT-PCR.Then highly associated genes in the early stage were determined also according to the ratio in mRNA,and seclected some of them analyzed their corresponding protein expression using Western blot.(2)Highly related genes in early stage of ESCC further detected in adjacent carcinoma tissues and distant non carcinoma tissues among 16 cases of ESCC by semi-quantitative RT-PCR.Normal esophageal tissues were used as control group,using immunohistochemistry detected them in pathological tissue sections of adjacent carcinoma tissues among 18 cases of ESCC.(3)Highly associated genes of early ESCC in Kazak were carried on topology analysis using STRING 9.0 online database and Pajek software.And 20 proteins were reported to demonstrate abnormal expression levels in the early stage of ESCC in other national region also had be done.Then analyzed the difference between them.Results:(1)36 highly related genes of ESCC in Kazak were selected by the ratio in mRNA levels comparation of ESCC tissues and distant non carcinoma tissues among 42 candidate genes.(2)Semi-quantitative RT-PCR and Western blot were used to detect 36 highly related genes in adjacent carcinoma tissues and distant non carcinoma tissues among 10 cases,it showed that 26 gengs were highly related to ESCC in early stage.(3)These differential expressed genes were confirmed at mRNA and protein levels,using RT-PCR in adjacent carcinoma tissues and distant non carcinoma tissues among 16 cases and immunohistochemistry in pathological tissue sections of adjacent carcinoma tissues among 18 cases,respectively.(4)26 genes expression profiling in early specimens showed that about 40 percent of individuals displayed genes expression combinations of survivin,ETV5,MMP-7,TIMP-1,MMP-2,c-myc,c-fos,MTA1,CDK4,cyclinD1,MCM4 and SKP2.(5)Two topology map were quite different that constructed by two group genes,one was established in this study and the other was reported in literature.And the map of node protein were also different.Conclusions:(1)c-fos,Ki67,MCM4,ETV5,SKP2,TIMP-1,MMP-2,CDK4,MEMD,MTA1,MMP-7,c-myc,c-jun,Survivin,CyclinD1,BAG1,Sp-17,c-erbB2,EphB4,CK-1,MGMT,periplakin,Snail,p33,Bax and E-cadherin in total 26 genes were highly associated with early ESCC in Kazak in Xinjiang.(2)Applying multiply genetic detection could contribute to diagnosis specificity,andgenes expression profiling might provide a laboratorial evidence for the clinical diagnosis of ESCC in early stage.(3)The topology map displayed national difference constructed by 26 highly related genes in early stage of ESCC in Kazak.These node proteins may helpful to explore the mechanism of ESCC.