The Relationship Between Tumor Related Gene Mutation And Leukemia Incident Based On Next-generation Sequencing

Objective: In this study,we used next-generation sequencing method to dig out the gene mutation spectrum of leukemia patients in Hainan area from the perspective of the tumor related target genes sequencing.It will helpful for the early diagnosis and prognosis of leukemia patients,and also helpful for study the pathogenesis and to find new therapeutic targets to guide the clinical treatment.The results of this study can also be used to detect risk genes in normal people,thus is conducive to discovery of potential factors.Methods: 104 cases of leukemia in Hainan General Hospital from August 2015 to December 2016 were recruited in this study,including 30 cases of acute myeloid leukemia,19 cases of acute lymphoblastic leukemia,19 cases of acute promyelocytic leukemia,28 cases of chronic myelogenous leukemia,and 8 cases of chronic lymphocytic leukemia.Through literature retrieval we get all the known tumor genes that associated with cancer.The target sequence capture array and Illumina Hiseq 2000 high-throughput sequencing technology were used for all exons of 605 target genes in sequencing,data analysis and Massarray verification.By means of high-throughput sample typing to establish all SNV Loci Library,and using SNV Loci Library information to extract the original sequencing results of each sample,then judging the result of the sample typing.Utilizing bi-directional sequencing balance chi-square test,binomial distribution and Bayesian method of probability estimation to genotype of candidate SNV site,and show the level of accuracy,further via Hardy-Wenberg equilibrium test evaluate the reliability of SNV site.We tried to detect mutations in 605 genes of the 104 samples and verify it with one generation of sequencing.Results: In this study,we identified 605 tumor related target genes closely related to cancer,104 patients leukemia patients were sequenced and verified,we found that:(1).The 2089 sites of these 605 genes have been mutated.(2).We found the gene mutation spectrum and target genes related to leukemia in Hainan,and integrated 50 genes into a test panel that has a large relationship with leukemia,such as,KMT2C?PDE4DIP?FBXW7?FANCD2 and USP6 etc.(3).The highest frequency of mutation was found to be KMT2 C.The relationship between the mutation and the clinical phenotype of AML(age,initial white cell count,CR rate)was confirmed,and it was found in patients younger than 50 years old and associated with higher initial white cell counts.The study confirmed that the rate of CR in the patients with KMT2 C mutation was low relatively,suggesting that the prognosis was maybe poor.(4).PDE4 DIP was no report on leukemia,but it had been found a high frequency mutation in this study.There was no statistical difference between different types of leukemia,and this gene mutation did not exist in the normal population.Therefore,it works on the leukemia needs to be further studied.Conclusions:(1).Preliminary study on gene mutation spectrum of leukemia patients in Hainan.The mutation frequency was found in the top 50 susceptibility genes,and the detection of panel was established.(2).The relationship between KMT2 C mutation and AML partial clinical phenotype is verified,may be associated with the development of leukemia(3).that offers a firm foundation for the establishment of standardized individual treatment and testing system in Hainan(4).It provides a theoretical basis for the early diagnosis of leukemia,and can identify genes related to the prognosis of the patients,so as to provide data support for clinical individual treatment.