Abstract: Objective To investigate the genetic information of endometrial cancer population in Huai'an area. Methods A total of 15 patients with endometrial cancer who underwent surgery in Huai'an Maternal and Child Health Hospital were enrolled and 5 ml of venous blood was collected from the patients. Then, 110 gene loci were detected by second-generation gene sequencing, and the gene mutation information of the patients was analyzed. Results There were 75 gene mutations in these patients. According to TGCA database analysis, 21 sense mutations were found, where ATM, MSH2, PIK3R1, PTEN, TET2 and TSC1 were tumor suppressor genes and ALK, CTNNB1, ERBB2, FGFR2, HNF1A, KIT, MTOR, PDGFRA, PPP2R1A and SF3B1 were oncogenes. The mutation rate of PTEN was the highest (46.67%), followed by PIK3R1, AKT2 andFOXO1, which were related to the pathogenesis of endometrial cancer. Conclusions Second-generation gene sequencing can determine the genetic mutation of endometrial cancer andexplore the possible molecular mechanism, which provide theoretical evidence for clinical development of new therapeutic targets.