Application Of Proteomics In Early Esophageal/Cardiac Cancer And Precancerous Lesions By MALDI-TOF-MS In The High Incidence Area

Objective:The south Taihang Mountain region of Hebei province is recognized as the highest incidence area for upper gastrointestinal cancer in the world.The morbidity and mortality rates of70-80%for all malignant tumors were attributed to esophageal cancer,gastric cardia adenocarcinoma and gastric carcinoma. Study reported that early detection, diagnosis and treatment is the effective way to improve survival rate.The result demonstrated endoscope screening is the best method for detection early upper gastrointestinal cancer and precancerous lesions in the high-risk region.endoscope screening not only need large human and material resources,but also is highly dependent on examiners'skills,so that endoscopy is not suitable for Large scale screening in population. It is imported issue to find a high-throughput and high sensitivity way for screening in the high-risk area.Cancer is a complex disease involving Multiple genes,multistage process, internal and external environment Interaction changing. There are a lot of tumor-associated protein in tumor tissue and body fluid in the process of cancer development. We try to find a high specificity and sensitivity tumor markers for screening by means of observing changes of the protein in serum that will be reduce Endoscopy scale.Surface enhanced laser desorption/ionization time of flight mass spectrometry and matrix assisted laser desorption/ionization time of flight mass spectrometry are most advanced proteomics approaches in the word, that is able to detect various proteins changing expressed by gene in different stage of prognosis of tumor.In this study,1The goal of this study was to screen for protein biomarkers in serum for precancerous lesions of EC and GAC using MALDI-TOF MS, magnetic beads and pattern recognition software.2We aim to identify biomarker pattern that can be monitor tumor recurrence though the level of proteins in serum compared preoperative and postoperative of endoscopic treatment.3Research ExPasy database, analyze the relationship between specific proteins changing and the development of EC and GAC.Methods:The objective of this study is general population in the high-risk area which has never been screened ranged from40-69years old. All patients came from CiXian, SheXian which accepted the initial endoscope screening or came from The Fourth Hospital of Hebei Medical University which performed the endoscopy to examine between2010and2011. We Recruited254esophageal carcinoma and precancerous lesions subjects, Including53patients with Esophageal mild dysplasia and moderate dysplasia, EDYS I-II,92patients with Esophageal severe dysplasia and Esophageal carcinoma in situ, EDYS Ⅲ/CIS),53patients with Advanced esophageal carcinoma, AEC),53patients with low-grade intraepithelial neoplasia,LGIN),89patients with high-grade intra epithelial neoplasia-intramucosal carcinoma, HGIN-IMC),60patients with invasive gastric cardia adenocarcinoma,57normal control NOR besides12cases of EDYS Ⅲ/CIS after endoscopic treatment,21cases of HGIN-IMC after endoscopic treatment. All patients were histopathologically confirmed by two specialists in the The Fourth Hospital of Hebei Medical University. All serum samples were pretreated with weak cation exchange (WCX) magnetic beads,and MALDI-TOF-MS Analyses,Peak detection was performed with Biomarker Wizard software3.21, After the assignment of biomarker candidates,Statistical analysis was performed using SPSS16.0software, finally1Construction of boosting decision tree for AEC/GCA and precancerous lesions.2Comparison of serum protein spectra between preoperative and postoperative of endoscopic treatment.3.Research ExPasy database.Results:1A total of146protein peaks were detected in EDYS Ⅰ-Ⅱ,EDYS Ⅲ/CIS,AEC and NOR cases in the training set,14protein peaks were selected to construct for the decision tree classification.This decision algorithm correctly classified90.91%of normal control,100%of EDYS Ⅰ-Ⅱ,89.66%of EDYSⅢⅢ /CIS, and81.26%of AEC. The peaks of2023,1307and1234were speculated zinc alpha2glycoprotein, Fibrinogen alpha chain and Glycosyltransferase like protein LARGE2by ExPasy database Retrieval,others were unknown.26protein peaks were significantly differently, when we compared the spectra of the12preoperative and postoperative sera obtained from each EDYSⅢ/CIS patient.the1563,1590peaks intensities were significantly increased in the preoperative sera. And the expression level of the this peaks was decreased after Endoscopic treatment. In addition The8108,8632,8960,32023peaks intensities were higher compare with preoperative sera.32023,8960were speculated zinc alpha2glycoprotein, Apolipoprotein All by ExPasy database.3A total of193protein peaks were detected in LGIN,HGIN-IMC, IGCA and NOR cases in the training set,15protein peaks were selected to construct for the decision tree classification.This decision algorithm correctly classified100%of normal control,100%of LGIN,85.246%of HGIN-IMC, and93.939%%of IGCA. The peaks of33313,8712and7992were speculated Leucine-rich a2glycoprotein(LRG),fragments of C3a and platelet factor4(PF4) by ExPasy database Retrieval,others were unknown.412protein peaks were significantly differently, when we compared the spectra of the21preoperative and postoperative sera obtained from each HGIN-IMC patient.the1951,1595,1771and2332peaks intensities were significantly increased in the preoperative sera compared with normal. And the expression level of the this peaks was decreased after Endoscopic treatment. In addition The31069,10289,9317,7992,7786,5918,4103,4660peaks intensities were higher compared with preoperative sera after operative (p<0.05).1771,2332,4103,5918,7786,9317and10289.51were speculated Apolipoprotein AIV, transgelin4103Inter-alpha-trypsin inhibitor heavy chain4Fibrinogen alpha chain,platelet factor4(PF4) Vitronectin V10subunit and10kDa chaperonin by ExPasy database.Conclusions:1The diagnostic pattern were Established by Differently protein peaks could accurately recognize EDYS Ⅰ-Ⅱ,EDYS Ⅲ/CIS,AEC patients and normal controls application of MALDI-TOF MS combined with magnetic beads and BPS.26protein peaks were significantly differently, when we compared the spectra of pre-and postoperative sera obtained from each EDYS Ⅲ/CIS patient.that was verify the changes of the difference in serum protein peak.3The diagnostic pattern were Established by Differently protein peaks could accurately recognize LGIN,HGIN-IMC,GCA patients and normal controls application of MALDI-TOF MS combined with magnetic beads and BPS.412protein peaks were significantly differently, when we compared the spectra of pre-and postoperative sera obtained from each HGIN-IMC patient.that was verify the changes of the difference in serum protein peak.5We speculated that the occurrence of esophageal/gastric cardia began in the EDYS Ⅰ-Ⅱ/LGIN or much earlier according the result of our studies.