Association Between Single Nucleotide Polymorphism And Sudden Decrease Of Heart Rate During Abdominal Lavage In Patients With Gastric Cancer

Objective:To identify single nucleotide polymorphisms(SNPs)associated with heart rate plunge during peritoneal lavage in patients with gastric cancer,and to investigate the association between SNPs and heart rate sudden drops.Methods:(1)Patients with gastric cancer who underwent gastrectomy in the Third Affiliated Hospital of Soochow University from March 2018 to March 2019 were selected as subjects.After gastrectomy and lymphadenectomy for all patients,500mL distilled water at 42℃ was injected into the abdominal cavity at the xiphoid process within 10cm of the abdominal wall within 5 seconds,and the lavage time was 15min.Heart rate(HR),Blood pressure(BP),Blood oxygen saturation(SaO2)and Bispectral Index(BIS)were recorded immediately before and after lavage.Heart rate change rate=[(heart rate before lavage-heart rate limit value after lavage)/heart rate before lavage]× 100%.(2)According to the change of heart rate,6 patients were randomly selected from the two groups with heart rate change>50%and<10%for whole exon sequencing(WES)analysis.Peripheral blood(2mL)was obtained from ulnar vein before surgery and DNA was extracted after purification,the library was constructed,the exon region was captured,and high-throughput sequencing was performed by Illumina HiSeq 2500 sequencer.Alignment(using BWAMEM to align the sequence with the standard genome),duplication removal and filtering,recombination and correction(using GATK to re-align the insertion and deletion of bases,and re-correct the bases),and detection(using GATK to analyze the SNP and Indel of samples).After sequencing analysis,the differential SNP sites were amplified by polymerase chain reaction(PCR)and verified by Sanger sequencing method.(3)Considering the possible influence of SNP in exon region on phenotype,the allele frequency,genotype frequency and haplotype of SNP in exon region with significant difference on heart rate during peritoneal lavage were analyzed in different groups(30%and 15%heart rate sudden drop were used as cut points respectively).And a simple detection method is establish,and three mutation sites of RECQL5(rs820196),TEP1(rs938886)and TEP1(rs1713449),gene are detected in a single tube based on a two-dimensional PCR technology.(4)Using the public database,our research group further supplemented and analyzed the genes related to heart rate sudden drop,downloaded the expression profile data of TEP1 and RECQL5 mutant group and non-mutant group from the database,carried out difference analysis,and enriched the differentially expressed genes in function and pathway,so as to provide a basis for further research on heart rate sudden drop.Thereby revealing the pathways possibly involved by the mutations of TEP1 and RECQL5 and the key core genes affected by the mutations;Inquire the position of amino acid change and the change of amino acid after the mutation of the three sites of the corresponding gene.Results:(1)A total of 194 patients were included in this study,including 138 males and 56 females.(2)The changes of heart rate in all patients during peritoneal lavage with warm distilled water were as follows:The heart rate decreased by 0%～10%in 65 cases,10～15%in 29 cases,15～30%in 50 cases,30～50%in 12 cases,50～100%in 4 cases,Six had cardiac arrest and 28 had elevated heart rates.(3)Compared with the reference genome,there were 4986 specific SNPs and 258 specific Indel in the group of heart rate changes>50%,there were 22294 specific SNPs and 1065 specific Indel in the group of heart rate changes<10%.The total number of SNPs and Indel in two group of patients was 16192 and 700 respectively.(4)Whole exon sequencing(WES)and polymerase chain reaction(PCR)confirmed that TTC5(rs74596138)in the non-coding region,TEP1(rs938886),TEP1(rs1713449),RECQL5(rs820196)in the exon region and TEP1-0948(rs1760909,rs1713448)in the intron region were associated with decreased heart rate.(5)There was no statistically significant difference in allele frequency and genotype frequency between TEP1(rs938886),TEP1(RS1713449)and RECQL5(rs820196).(6)The linkage unbalance test showed that there was a strong linkage unbalance between TEP1(rs938886)and TEP1(rs1713449).(7)For RECQL5(rs820196),patients carrying the TC genotype were associated with a significantly increased risk of adverse heart rate decline compared with individuals carrying the TT genotype.(8)Compared with the wild type GG-CC-TT of three SNP loci TEP1(rs938886),TEP1(rs1713449)and RECQL5(rs820196),the risk of heart rate decrease in gastric cancer patients carrying GC-CT-TC genotype was significantly increased.(9)In the interaction analysis of environmental factors,smoking and hypertension may have potential interaction with the decrease of heart rate in patients with gastric cancer.(10)The Mutations of TEP1(rs938886),TEP1(rs1713449)and RECQL5(rs820196)were all harmless to the tumor,but they had some effects on the heart rate changes of gastric cancer patients during peritoneal lavage.In particular,RECQL5(rs820196)was significantly associated with heart rate sudden drop during peritoneal lavage in patients with gastric cancer.Conclusion:(1)Abdominal lavage can cause heart rate changes in patients with gastric cancer;(2)Mutant heterozygote TC of RECQL5(rs820196)increases the adverse risk of heart rate sudden drop.(3)Compared with the wild type GG-CC-TT of three SNP loci TEP1(rs938886),TEP1(rs1713449)and RECQL5(rs820196),the risk of heart rate decrease in gastric cancer patients with GC-CT-TC genotype was significantly increased.(4)There may be a potential interaction between smoking,hypertension and heart rate decline in patients with gastric cancer.(5)TEP1(rs938886),TEP1(rs1713449)and RECQL5(rs820196)could affect the heart rate changes of gastric cancer patients during peritoneal lavage to some extent,especially RECQL5(rs820196)was significantly associated with the heart rate changes of gastric cancer patients during peritoneal lavage.