AIM: To evaluate the effect of promoter region polymorphisms of toll-like receptor(TLR)2-196 to-174 del and TLR4-1607T/C(rs10759932) on m RNA and protein expression in tumor tissue and of TLR4+896A/G(rs4986790) on col...AIM: To evaluate the effect of promoter region polymorphisms of toll-like receptor(TLR)2-196 to-174 del and TLR4-1607T/C(rs10759932) on m RNA and protein expression in tumor tissue and of TLR4+896A/G(rs4986790) on colorectal cancer(CRC) risk.METHODS: The TLR2-196 to-174 del polymorphism was investigated using allele-specific polymerase chain reaction(PCR) and the TLR4-1607T/C and TLR4+896A/G by PCR-restriction fragment length p o l y m o r p h i s m( R F L P). W e g e n o t y p e d 4 3 4 D N A samples from 194 CRC patients and 240 healthy individuals. The m RNA relative quantification(RQ) was performed in 40 tumor tissue samples by quantitative PCR Taq Man assay, using specific probes for TLR2 and TLR4 genes, and ACTB and GAPDH reference geneswere used as endogenous controls. Protein expression was analyzed by immunohistochemistry with specific primary antibodies.RESULTS: No association was found for TLR4-1607T/C and TLR4+896A/G by three statistical models(logadditive, dominant and recessive). However, based on dominant and log-additive models, the polymorphic variant TLR2-196 to-174 del was associated with increased CRC risk [dominant: odds ratio(OR) = 1.72, 95%CI: 1.03-2.89; P = 0.038 and log-additive: OR =1.59, 95%CI: 1.02-2.48; P = 0.039]. TLR2 m RNA expression was increased in tumor tissue(RQ = 2.36) when compared to adjacent normal tissue(RQ = 1; P < 0.0001), whereas the TLR4 m RNA showed a basal expression(RQ = 0.74 vs RQ = 1, P = 0.452). Immunohistochemistry analysis of TLR2 and TLR4 protein expression was concordant with the findings of m RNA expression. In addition, the TLR2-196 to-174 del variant carriers showed m RNA relative expression 2.19 times higher than wild-genotype carriers. The TLR2 protein expression was also higher for the TLR2-196 to-174 del variant carriers [117 ± 10 arbitrary unit(a.u.) vs 95 ± 4 a.u., P = 0.03]. However, for the TLR4-1607T/C polymorphism no significant difference was found for both m RNA(P = 0.56) and protein expression(P = 0.26).CONCLUSION: Our findings suggest that TLR2-196 to-174 del polymorphism increases TLR2 m RNA expression and is associated with higher CRC risk, indicating an important role in CRC genetic susceptibility.展开更多
AIM To evaluated the association of the risk factors and polymorphisms in MTHFR C677 T,MTHFR A1298 C,MTR A2756 G and MTRR A66 G genes.METHODS Patients with cirrhosis(n=116),hepatocellular carcinoma(HCC)(n=71)and contr...AIM To evaluated the association of the risk factors and polymorphisms in MTHFR C677 T,MTHFR A1298 C,MTR A2756 G and MTRR A66 G genes.METHODS Patients with cirrhosis(n=116),hepatocellular carcinoma(HCC)(n=71)and controls(n=356)were included.Polymerase chain reaction followed by enzymatic digestion and allelic discrimination technique real-time PCR techniques were used for analysis.MINITAB-14.0and SNPstats were utilized for statistical analysis.RESULTS Showed that age≥46 years(OR=10.31;95%CI:5.66-18.76;P<0.001)and smoking(OR=0.47;95%CI:0.28-0.78;P=0.003)were associated with cirrhosis.Age≥46 years(OR=16.36;95%CI:6.68-40.05;P<0.001)and alcohol habit(OR=2.01;95%CI:1.03-3.89;P=0.039)were associated with HCC.MTHFR A1298 C in codominant model(OR=3.37;95%CI:1.52-7.50;P=0.014),recessive model(OR=3.04;95%CI:1.43-6.47;P=0.0051)and additive model(OR=1.71;95%CI:1.16-2.52;P=0.0072)was associated with HCC,as well as MTR A2756 G in the additive model(OR=1.68;95%CI:1.01-2.77;P=0.047),and MTRR A66 G in the codominant model(OR=3.26;95%CI:1.54-6.87;P<0.001),dominant model(OR=2.55;95%CI:1.24-5.25;P=0.007)and overdominant model(OR=3.05;95%CI:1.66-5.62;P<0.001).MTR A2756 G in the additive model(OR=1.54;95%CI:1.02-2.33;P=0.042)and smokers who presented at least one polymorphic allele for MTRR A66G(OR=1.71;95%CI:0.77-3.82;P=0.0051)showed increased risk for cirrhosis.There was no association between clinical parameters and polymorphisms.CONCLUSION Age≥46 years,alcohol habit and MTR A2756 G,MTHFR A1298 C and MTRR A66 G polymorphisms are associated with an increased risk of HCC development;age≥46 years,tobacco habit and the MTR A2756 G polymorphism are associated with cirrhosis.展开更多
基金Supported by Grants from Brazilian agencies FAPESP,No.2012/15036-8and CNPq,No.304870/2012-9
文摘AIM: To evaluate the effect of promoter region polymorphisms of toll-like receptor(TLR)2-196 to-174 del and TLR4-1607T/C(rs10759932) on m RNA and protein expression in tumor tissue and of TLR4+896A/G(rs4986790) on colorectal cancer(CRC) risk.METHODS: The TLR2-196 to-174 del polymorphism was investigated using allele-specific polymerase chain reaction(PCR) and the TLR4-1607T/C and TLR4+896A/G by PCR-restriction fragment length p o l y m o r p h i s m( R F L P). W e g e n o t y p e d 4 3 4 D N A samples from 194 CRC patients and 240 healthy individuals. The m RNA relative quantification(RQ) was performed in 40 tumor tissue samples by quantitative PCR Taq Man assay, using specific probes for TLR2 and TLR4 genes, and ACTB and GAPDH reference geneswere used as endogenous controls. Protein expression was analyzed by immunohistochemistry with specific primary antibodies.RESULTS: No association was found for TLR4-1607T/C and TLR4+896A/G by three statistical models(logadditive, dominant and recessive). However, based on dominant and log-additive models, the polymorphic variant TLR2-196 to-174 del was associated with increased CRC risk [dominant: odds ratio(OR) = 1.72, 95%CI: 1.03-2.89; P = 0.038 and log-additive: OR =1.59, 95%CI: 1.02-2.48; P = 0.039]. TLR2 m RNA expression was increased in tumor tissue(RQ = 2.36) when compared to adjacent normal tissue(RQ = 1; P < 0.0001), whereas the TLR4 m RNA showed a basal expression(RQ = 0.74 vs RQ = 1, P = 0.452). Immunohistochemistry analysis of TLR2 and TLR4 protein expression was concordant with the findings of m RNA expression. In addition, the TLR2-196 to-174 del variant carriers showed m RNA relative expression 2.19 times higher than wild-genotype carriers. The TLR2 protein expression was also higher for the TLR2-196 to-174 del variant carriers [117 ± 10 arbitrary unit(a.u.) vs 95 ± 4 a.u., P = 0.03]. However, for the TLR4-1607T/C polymorphism no significant difference was found for both m RNA(P = 0.56) and protein expression(P = 0.26).CONCLUSION: Our findings suggest that TLR2-196 to-174 del polymorphism increases TLR2 m RNA expression and is associated with higher CRC risk, indicating an important role in CRC genetic susceptibility.
基金the Faculdade de Medicina de Sao Jose do Rio Preto,FAMERP and Medical School FoundationFUNFARME for their institutional supportUPGEM-Genetics and Molecular Biology Research Unit
文摘AIM To evaluated the association of the risk factors and polymorphisms in MTHFR C677 T,MTHFR A1298 C,MTR A2756 G and MTRR A66 G genes.METHODS Patients with cirrhosis(n=116),hepatocellular carcinoma(HCC)(n=71)and controls(n=356)were included.Polymerase chain reaction followed by enzymatic digestion and allelic discrimination technique real-time PCR techniques were used for analysis.MINITAB-14.0and SNPstats were utilized for statistical analysis.RESULTS Showed that age≥46 years(OR=10.31;95%CI:5.66-18.76;P<0.001)and smoking(OR=0.47;95%CI:0.28-0.78;P=0.003)were associated with cirrhosis.Age≥46 years(OR=16.36;95%CI:6.68-40.05;P<0.001)and alcohol habit(OR=2.01;95%CI:1.03-3.89;P=0.039)were associated with HCC.MTHFR A1298 C in codominant model(OR=3.37;95%CI:1.52-7.50;P=0.014),recessive model(OR=3.04;95%CI:1.43-6.47;P=0.0051)and additive model(OR=1.71;95%CI:1.16-2.52;P=0.0072)was associated with HCC,as well as MTR A2756 G in the additive model(OR=1.68;95%CI:1.01-2.77;P=0.047),and MTRR A66 G in the codominant model(OR=3.26;95%CI:1.54-6.87;P<0.001),dominant model(OR=2.55;95%CI:1.24-5.25;P=0.007)and overdominant model(OR=3.05;95%CI:1.66-5.62;P<0.001).MTR A2756 G in the additive model(OR=1.54;95%CI:1.02-2.33;P=0.042)and smokers who presented at least one polymorphic allele for MTRR A66G(OR=1.71;95%CI:0.77-3.82;P=0.0051)showed increased risk for cirrhosis.There was no association between clinical parameters and polymorphisms.CONCLUSION Age≥46 years,alcohol habit and MTR A2756 G,MTHFR A1298 C and MTRR A66 G polymorphisms are associated with an increased risk of HCC development;age≥46 years,tobacco habit and the MTR A2756 G polymorphism are associated with cirrhosis.
