Mplification of gene fragmentsPolymerase chain reaction (PCR) was carried out by using the Bio-Rad CFX 96 real-time PCR method (Bio-Rad, USA). We established a PCR reaction system (20 l): Taq PCR Master Mix (2X) 10ul; 10p mol/ul Forward primer 0.4ul; 10p mol/ul 5-HT6 Receptor Modulator web Reverse primer 0.4ul; Template DNA 2ul; ddH2O 7.6ul; The reaction conditions: pre-denaturation at 95 for 3min, denaturation atPLOS 1 | https://doi.org/10.1371/journal.pone.0253474 June 30,three /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 10 min.Statistical analysisThe allele and genotype frequencies were calculated straight. SPSS20.0 software program was employed for statistical analysis. Measurement information were expressed as imply tandard deviation ( sd). x The odds ratio (OR) and 95 Self-assurance Interval (95 CI) represented the gene polymorphism internet sites and also the susceptibility threat of colorectal cancer. Hardy-Weinberg equilibrium was analyzed by 2test. The T-type calcium channel custom synthesis cut-off worth of substantial distinction was P0.05.Results1. By way of the genetic association analysis of 168 CRC instances and 710 controls, a significant association in between CYP24A1 polymorphism carriers (rs6013905AX and rs2762939GX) and CRC (p0.05) (Tables 1 and two) was identified. Compared together with the control group, CRC sufferers 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 substantially increased incidence danger. Also, the frequency with the rs6013905 A allele was related with an elevated incidence risk of CRC (P = 0.03, OR = 1.32, 95 CI: 1.03.69). The rs6013905 polymorphism had a considerable association with CRC within the dominant model (P = 0.02, OR = 1.89, 95 CI: 1.09.29). Compared with the handle group, CRC sufferers who carried the rs2762939 GC genotype were drastically at a higher incidence danger (P = five.56 ten, OR = 1.63, 95 CI: 1.15.31). Carriers of rs2762939 (GX) genotype have been also significantly connected with an elevated threat of CRC (P = 0.02, OR = 1.52, 95 CI: 1.08.13) in the dominant model. For CRC inside the JiamusiTable 1. Qualities from the study population. Variable Age (years) 60 60 Mean SD Sex Male Female Location with the principal tumor Colon Rectum Basic classification of tumors Ulcerative variety Uplift variety Degree of differentiation Low Higher 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 Control No. ( ) 206 N.S. P-valuePLOS A single | https://doi.org/10.1371/journal.pone.0253474 June 30,4 /PLOS ONECYP24A1 gene polymorphism with colorectal cancerTable two. Comparative evaluation of CYP24A1 polymorphisms amongst the CRC and manage 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.
