Mplification of gene fragmentsPolymerase chain reaction (PCR) was carried out by utilizing the Bio-Rad CFX 96 PRMT1 Biological Activity real-time PCR system (Bio-Rad, USA). We established a PCR reaction method (20 l): Taq PCR Master Mix (2X) 10ul; 10p mol/ul Forward primer 0.4ul; 10p mol/ul Reverse primer 0.4ul; Template DNA 2ul; ddH2O 7.6ul; The reaction conditions: pre-denaturation at 95 for 3min, denaturation atPLOS A single | https://doi.org/10.1371/journal.pone.0253474 June 30,3 /PLOS ONECYP24A1 gene polymorphism with colorectal cancer95 for 30s, annealing temperature of 25s at Tm, elongation at 72 for 30s, duplicate 36 cycles, terminal elongation at 72 for ten min.Statistical analysisThe allele and genotype frequencies had been calculated straight. STAT6 medchemexpress SPSS20.0 computer software was utilised for statistical evaluation. Measurement data had been expressed as imply tandard deviation ( sd). x The odds ratio (OR) and 95 Self-confidence Interval (95 CI) represented the gene polymorphism web-sites and also the susceptibility risk of colorectal cancer. Hardy-Weinberg equilibrium was analyzed by 2test. The cut-off worth of important difference was P0.05.Results1. By means of the genetic association evaluation of 168 CRC cases and 710 controls, a important association among CYP24A1 polymorphism carriers (rs6013905AX and rs2762939GX) and CRC (p0.05) (Tables 1 and 2) was identified. Compared using the manage group, CRC individuals carrying rs6013905 GA genotype (P = 0.04, OR = 1.79, 95 CI: 1.01.19) and AA genotype (P = 0.02, OR = 2.02, 95 CI 1.13.63) had a considerably elevated incidence threat. Also, the frequency from the rs6013905 A allele was associated with an improved incidence danger of CRC (P = 0.03, OR = 1.32, 95 CI: 1.03.69). The rs6013905 polymorphism had a substantial association with CRC in the dominant model (P = 0.02, OR = 1.89, 95 CI: 1.09.29). Compared with all the control group, CRC sufferers who carried the rs2762939 GC genotype have been considerably at a higher incidence danger (P = 5.56 10, OR = 1.63, 95 CI: 1.15.31). Carriers of rs2762939 (GX) genotype had been also drastically related with an elevated risk of CRC (P = 0.02, OR = 1.52, 95 CI: 1.08.13) in the dominant model. For CRC inside the JiamusiTable 1. Qualities of the study population. Variable Age (years) 60 60 Imply SD Sex Male Female Location on the primary tumor Colon Rectum Common classification of tumors Ulcerative variety Uplift sort Degree of differentiation Low High TNM I I III N.S. represent no substantial. https://doi.org/10.1371/journal.pone.0253474.t001 83(49.40) 81(48.21) 153 (91.07) 9 (five.35) 130 (77.38) 34 (20.24) 96 (57.14) 72 (42.85) 98 (58.33) 70 (41.67) 112 (54.36) 94 (45.63) 63 (37.50) 105 (62.50) 62.990.91 97 (47.09) 109(52.91) 59.424.99 N.S. Case No. ( ) 168 Manage No. ( ) 206 N.S. P-valuePLOS One | https://doi.org/10.1371/journal.pone.0253474 June 30,four /PLOS ONECYP24A1 gene polymorphism with colorectal cancerTable two. Comparative analysis of CYP24A1 polymorphisms involving the CRC and control groups. Genotype rs6013905 GG GA AA Dominant model (AX) Recessive model (AA) G A rs2762939 CC CG GG Dominant model (GX) Recessive model (GG) C G rs6068816 GG GA AA Dominant model (AX) Recessive model (AA) G A https://doi.org/10.1371/journal.pone.0253474.t002 69(0.41) 81(0.48) 18(0.11) 99(0.59) 18(0.11) 219(0.65) 117(0.35) 261(0.37) 341(0.48) 108(0.15) 449(0.63) 108(0.15) 863(0.61) 557(0.39) Ref. N.S. N.S. N.S. N.S. Ref. N.S. 77(0.46) 83(0.49) 8(0.05) 91(0.54) eight(0.05) 237(0.71) 99(0.29) 399(0.56) 264(0.37) 47(0.07) 311(0.44) 47(0.07) 1062(0.