| ?Objective?Based on Affymeytrix Axiom chip and bioinformatics method,we constructed the interaction network of the regulatory gene of autophagy related pathway to screen the key gene and signal transduction pathway in gastric cancer.By comparing and analyzing the susceptibility genes and genetic markers associated with autophagy pathway in gastric cancer patients and controls,we studied the relationship between the susceptibility genes and the risk of gastric cancer.To explore its influence on the risk of gastric cancer,we constructed the best interaction model of genes and genes(environment),which provided the basis for the prevention and clinical study of gastric cancer,and provided a reference for the further study of the pathogenesis of gastric cancer.?Methods?1.Using 920K Axiom Precision Medicine Research Array produced by Affymetrix,we detected genotypes of single nucleotide polymorphism(SNP)of the total DNA in peripheral blood leukocytes from 96 patients with gastric adenocarcinoma and 96healthy controls from Xianyou,Fujian Province.The key genes and signal transduction pathway of autophagy in gastric cancer were screened by using SPSS20.0 software to carry out the analysis of chi-square trend test and combining with bioinformatics(KEGG PATHWAY website,Gene Ontology database,STRING website and so on).2.To further screen the genes and their SNPs related to the formation pathway of autophagosome,we increased the sample size of 622 cases in the case group and the control group respectively.Using the technique platform of Sequenom Mass ARRAY,we detected the genotypes of 17 SNP of 11 genes(AKT1,ATG2B,CAMK2,DAPK1,EIF2AK4,IRS1,PIK3CD,PIK3R1,PRKACA,UVRAG,ZFYVE1).And we analyze the association of SNP of each gene with gastric cancer by Cox regression.3.Combining significant environmental factors in previous studies and SNP,GMDR software was used to construct the best interaction model of gene-gene and gene-environment.Logistic regression analysis,crossover analysis method and EXCEL table developed by Andersson et al were used to analyze the relationship between the best interaction model and gastric cancer at different sites.The OR value and 95%confidence limit were calculated.?Results?1.SNP chip data showed that the chip contained more than 920000 SNPs,including 3702 SNPs of genes related to autophagy pathway,which was analyzed by bioinformatic approaches.Gastric cancer population was involved in signal pathway such as mTOR signaling pathway,insulin resistance,AMPK signal pathway,PI3K-AKT signal pathway and other tumor-related signaling pathways.These signal pathways were closely related to the formation of autophagosome.The main nodal genes are KT1?ATG5?ATG7?ATG16L1?IRS1?mTOR?ZFYVE1?UVRAG?ULK2?ATG2B?EIF2K4and so on.2.Further analysis revealed that the Beclin1-Vps34-p150 signal pathway may be related to the occurrence of gastric cancer,according to the analysis of the autophagic related signaling pathway related to the occurrence of gastric cancer in previous studies.The patients with ZFYVE1 rs12882235(CC)?DAPK1 rs721936(AA+AG)?UVRAG rs11289696(DEL)had higher risk of gastric cancer and gastric cardia cancer than those without them.The adjusted OR were 2.394 and 5.767,respectively,and with the increase of the number of carrying bad genes,the risk of gastric cancer and cardiac cancer increased gradually.The association between 17 SNPs of 11 key node genes such as AKT1?ATG2B?CAMKK2?DAPK1?EIF2AK4?IRS1?PIK3CD?PIK3R1?PRKACA?UVRAG?ZFYVE1 in the corresponding pathway and gastric cancer were analyzed.The results showed that ATG2B rs10134160(T>C)?ZFYVE1 rs12882235(T>C)and IRS1rs10205233(T>C)reduced the risk of gastric cancer,which OR_a(95%CI)were0.770(0.635,0.935)?0.767(0.650,0.906)and 0.794(0.643,0.982).Rs12882235(T>C)and EIF2AK4 rs56143280(GG)reduced the risk of gastric cardia cancer,which OR_a(95%CI)were 0.724(0.573,0.916)and 0.702(0.496,0.992).Rs10134160(T>C)reduced the risk of gastric non-cardia cancer,which OR_a(95%CI)was 0.653(0.497,0.859).3.The results of gene-gene(environment)combination and interaction analysis showed that the best interaction model of gene was rs10134160,rs59513966 and rs60599616.The high risk exposure combination of gastric cancer was showed in individual who carried ATG2B rs59513966(CC),hard food diet and intaking pickled vegetables.The higher risk of gastric cancer was showed in individual who carried rs10134160(CC)and hard food diet(OR=5.568),and who carried rs10134160(CC)and intaking pickled vegetables(OR=1.845).The higher risk of gastric cancer was showed in individual who carried rs10205233(CC)and hard food diet(OR=5.613),and who carried rs10205233(CC)and intaking pickled vegetables(OR=1.734).The higher risk of gastric cancer was showed in individual who carried rs12882235(CC)and hard food diet(OR=6.550),and who carried rsrs12882235(CC)and intaking pickled vegetables(OR=2.401).The negative multiplicative and additive interaction were showed between the individual who carried rs12882235 and smoking,and 69.8%of the patients who reduced the risk of gastric cancer were attributed to the additive interaction.The higher risk of gastric cancer was showed in individual who carried rs56143280A allele and hard food diet(OR=5.607),and who carried rs56143280A allele and intaking pickled vegetables(OR=1.828).?Conclusion?1.The single nucleotide polymorphisms in autophagy associated AMPK signa pathway,mTOR signal pathway and PI3K-AKT signal pathway may be related to the genetic susceptibility of gastric cancer population in Xianyou County,Fujian Province,and these pathways were at the stage of autophagy vesicle formation.2.The susceptibility SNPs of gastric cancer in different parts were different.ATG2B rs10134160?ZFYVE1 rs12882235 and IRS1 rs10205233 mutation all reduced the risk of gastric cancer in the local population;rs12882235and EIF2AK4 rs56143280mutation reduced the risk of gastric cardia cancer;rs10134160 mutation reduced the risk of non-cardia cancer.Autophagy related Beclin1-Vps34-p150 signal pathway may play an important role in the gastric cancer.3.Rs10134160?rs12882235?rs10205233?rs56143280 were associated with hard food diet and intaking pickled vegetables,and it showed that there were multiplicative and additive interaction between rs12882235 and smoking.The risk of gastric cancer can be reduced by guiding individuals with bad genotypes to develop good habits,such as not smoking,drinking less,not eating hard food and so on. |
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