Abstract: Objective To analyze the characteristics of KRAS as well as other tumor-related gene mutation in colorectal cancer patients, and provide evidences for treatment and prognosis of these patients. Methods Totally 50 colorectal cancer patients with KRAS exon 2 G12/13 mutation admitted to our hospital from January 2015 to June 2016 were enrolled in this study. Next-generation sequencing (NGS) technology was used to detect the hot spot mutations of 59 tumor-related genes in FFPE samples. Results Totally 65 mutations in 12 genes were detected except KRAS. The abundance of mutations was over 10% in TP53 (62%), APC (46%), PIK3CA (22%) and SMAD4 (14%). The mutation frequency or hotspot distribution of these genes was associated with the growth of colorectal tumors. Conclusion TP53, APC, PIK3CA, SMAD4 may be related to KRAS gene and its pathway in colorectal cancer.