The Structure Analysis Of EML4-ALK Fusion Genelociand The Association Study Between-308 Signal Nucleotidepolymorphism(SNP) Of TNF-α Gene And Primary Lung Cancer

Objective:Lung cancer is one of the most prevalence malignances,of which non-small cell lung cancer(NSCLC)account for 80%~85%of lung cancer.About 25%~30%was terminally when they see a doctor,about 40%~50%for distant metastasis,have lose the best chance of surgical excision,chemotherapy and radiotherapy is the main treatment.But the effective of chemotherapy also is only 30%~40%,and many patients with vomiting,hair loss and other serious side effects,it is difficult to tolerate.Molecular targeted therapy is promising in treating some patients with NSCLC.EML4-ALK is a lung cancer-induced fusion gene,which was first discovered in Japan in 2007.Studies show that the positive rate of EML4-ALK is about 0.9%~2.6%in patients with non-small-cell lung cancer(NSCLC)by Inamura and Shinmura.Former researches indicated that EML4-ALK fusion and EGFR,K-ras mutation were excluded mutually.The EML4-ALK fusion oncogene represents a novel molecular target which appears mainly in lung adenocarcinoma.So far,the mechanism of EML4-ALK fusion gene is still unclear.The purpose of this study is based on H2228 cell lines EML4-ALK fusion gene loci to discuss the fused eml4-alk fusion method and the possible mechanism of integration and we also through to check the general population and primary lung cancer among patients with tumor necrosis factor alpha(TNF alpha)gene-308 site the single nucleotide polymorphism(SNP),evaluate the role of TNF-α-308G/A single nucleotide polymorphism(SNP)and the risk of primary lung cancer in Chinese population.Methods:This study is divided into two parts:the first part is using nested PCR to amplification the EML4-ALK fusion gene.Though NCBI-BIAST(National Center for Biotechnology Information)and CENSOR analysis fusion gene sequence structure and fusion mechanism were discussed.The second part is to address this study which a total of 250 patients and 447 healthy controls were recruited for.Genotyping were done by using Taq Man technology.Result:1.We got a 3800bp intro sequence.In the 3800bp intro sequence,we found three Alu repeat sequence ALU-sx,ALU1-TS,ALU-JO near the breakpoint region,and MIR sequence located near in ALK gene breakpoint.And two SNP are been found.2.Frequencies of(GG),(A/G)and(A/G+AA)genotypes of-308G/A SNP in TNF-αgene were 183(73.2%),67(26.8%)and 67(26.8%)in patients,and 406(90.8%),39(8.7%)and 41(9.2%)in controls respectively.The distribution of polymorphism frequencies in the case group and the control group were statistically significant for Chinese population(P<0.05).Conclusion:1.There are three ALU repeats and MIR sequence in the EML4-ALK fusion sequence.And some hot point sequence CTGT and CCAGC are found near the breakpoint.2.The results indicated that TNF-αgene polymorphism at position-308G/A appear to be associated with susceptibility to lung cancer in Chinese Han population.