Association Between Single Nucleotide Polymorphisms(SNPs) Of CRTC2,TGF?2 And Non-small Cell Lung Cancer(NSCLC)

Objective:To verify whether the two SNP sites of CRTC2,rs11264680 and rs8450,and the one SNP site of TGF?2,rs6658835 increase the risk of non-small cell lung cancer(NSCLC).To explore the association between genotype and allele frequency of rs11264680,rs8450 and rs6658835 mutation loci and gender,smoking history,tutor pathological type,lymph node status and clinical staging of patients with NSCLC.Methods:1.Specimen collection:Peripheral blood 3-4ml of elderly patients with non-small cell lung cancer diagnosed by the Department of Critical Care and Respiratory Medicine,Department of Thoracic Surgery was collected in The First Affiliated Hospital of Kunming Medical University,and 1-2ml peripheral blood of healthy people in the physical examination center was collected as the control;CRTC2,The number of NSCLC patients was 243,and the number of healthy control patients was 353.TGF ?2,The number of NSCLC patients was 272,and the number of healthy control patients was 338.2.DNA extraction,Beijing Tiangen 'blood genomic DNA extraction kit' was used to extract DNA according to the instructions,The A280/260 ratio and DNA concentration of the extracted part were determined by a microquantifier.3.Target fragment amplification,? Primer design,the forward primer and the reverse primer of rs11264680,rs8450 and rs6658835 were designed respectively by Primer3.?Polymerase chain reaction(PCR)amplification of the target product.? PCR product detection,agarose gel electrophoresis was used to detect the target fragment length.4.Mutation detection,detection of mutations by direct sequencing.5.Genotyping:by‘Chromas MFC Application' software.6. Statistical analysis:All the data were analyzed by the SPSS 22.0 statistical software.Alleles and genotype frequencies of the three SNP loci were obtained by sequencing,and the Hardy-Weinberg equilibrium was evaluated by Chi-square test;and then Pearson Chi-square test was used to compare the genotype frequency,allele frequency,dominant model,recessive model and overdominant model between the NSCLC and the control group.Stratified analysis was performed in NSCLC patient,including gender,smoking history,pathological type,lymph node status,and clinical stage were grouped,and the genotype and the allele frequency were compared between NSCLC and control groups.Results:The SNP sites of CRTC2(rs11264680 and rs8450)and TGF ? 2(rs6658835)were not correlated with NSCLC.There were no significant differences in genotype,allele frequency,dominant model,recessive model and overdominant model between NSCLC patients and the control group.Stratified analyzed indicates that rs 11264680,rs8450 and rs6658835 polymorphisms were not associated with the clinical characteristics of NSCLC patients.Conclusion:In the present study,SNP sites of CRTC2 and TGFP2 genes are not correlated with NSCLC susceptibility,and were not correlated with the clinical characteristics of NSCLC,including gender,smoking history,pathological type,lymph node status and clinical stage.CRTC2 and TGF 2 may not be a novel genetic marker of NSCLC.