目的观察艾灸对结肠炎相关性结肠癌(CAC)大鼠的干预作用,从嘌呤受体P2X7R与Wnt/b-catenin信号通路探讨可能的效应机制。方法将SD大鼠随机分为正常组、CAC组、隔药灸组、隔姜灸组。CAC组、隔药灸组、隔姜灸组均采用腹腔注射AOM联合DSS法...目的观察艾灸对结肠炎相关性结肠癌(CAC)大鼠的干预作用,从嘌呤受体P2X7R与Wnt/b-catenin信号通路探讨可能的效应机制。方法将SD大鼠随机分为正常组、CAC组、隔药灸组、隔姜灸组。CAC组、隔药灸组、隔姜灸组均采用腹腔注射AOM联合DSS法制备CAC大鼠模型,隔药灸与隔姜灸组均取天枢(双)、气海穴进行治疗。记录各组大鼠体质量、疾病活动指数(DAI)和成瘤率;通过HE染色观察艾灸对CAC大鼠结肠损伤的干预效应;通过RT-q PCR和Western Blot技术,检测艾灸对CAC大鼠结肠组织C-myc、Wnt1、b-catenin、GSK-3bm RNA和P2X7R蛋白表达的调节作用。结果与正常组相比,CAC组大鼠体质量显著降低、DAI增高、成瘤率明显增加(P<0.05),结肠组织可见腺管共壁背靠背和筛样结构,高级别腺癌形成。CAC组大鼠结肠组织P2X7R蛋白表达显著下调(P<0.05),C-myc、b-catenin、GSK-3b、Wnt1 m RNA的表达均显著上调(P<0.05)。与CAC组相比,隔药灸组和隔姜灸组大鼠体质量增加,DAI降低(P<0.05),结肠组织P2X7R蛋白表达显著上调,C-myc m RNA下调(P<0.05);隔姜灸组成瘤率明显降低(P<0.05),且Wnt1、b-catenin、GSK-3bm RNA表达显著下调(P<0.05)。结论隔药灸、隔姜灸均能调节CAC大鼠结肠组织P2X7R及C-myc的异常表达,且隔姜灸还能下调CAC大鼠结肠组织Wnt1、b-catenin、GSK-3bm RNA的表达。展开更多
Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells. The mechanisms underlying expression and regulation of Lin28A have been well documented, but such inf...Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells. The mechanisms underlying expression and regulation of Lin28A have been well documented, but such information for Lin28B is limited. In this study, a fragment of the Lin28B promoter was cloned, the pEGFP-pLin28B vector was constructed. DF-1 chicken fibroblasts were transfected and the expression of green fluorescent protein (GFP) was measured. Furtherly, Lin28B promoter of different lengths fragments was cloned using the chromosome-walking method and the fragments were ligated into the PGL3-Basic vector, and transfected into DF-1 cells. Results of dual-luciferase reporter assay showed that the core of the Lin28B promoter was included in the sequence from –1 431 to –1 034 bp. The binding sites of the transcription factor TCF7L2 was showed within this sequence by bioinformatics analysis. The promoter activity of Lin28B was downregulated (P<0.05) when the TCF7L2 binding site was mutated. Further experiments suggested that Lin28B promoter activity responded to the activation or inhibition of Wnt signaling. Results of chromatin immunoprecipitation and quantitative PCR showed that β-catenin-TCF7L2 may be enriched in the Lin28B promoter core area. In vivo and in vitro activation or inhibition of Wnt signaling significantly up- or down-regulated (P<0.05) Lin28B expression. H3K4me2 enriched in the promoter of Lin28B, which affected the regulation of Wnt signaling to Lin28B. In conclusion, our results showed that H3K4me2 and Wnt5a/β-catenin/TCF7L2 were the positive regulators of Lin28B expression. Findings of this study may lay a theoretical foundation for illuminating the mechanism underlying Lin28B expression.展开更多
AIM: To explore the association between TCF7L2 rs12255372 and rs7903146 single nucleotide polymorphisms(SNPs) and gastric cancer risk in Venezuelan patients.METHODS: We performed a case-control study including 122 par...AIM: To explore the association between TCF7L2 rs12255372 and rs7903146 single nucleotide polymorphisms(SNPs) and gastric cancer risk in Venezuelan patients.METHODS: We performed a case-control study including 122 paraffin-embedded archived intestinaltype gastric cancer samples and 129 biopsies obtained by superior endoscopy from chronic gastritis patients. Gastric cancer samples were classified according the degree of carcinoma differentiation. Genomic DNA was extracted from tissues, and the two SNPs of TCF7L2 gene(rs12255372 and rs7903146) were genotyped by polymerase chain reaction-restriction fragment length polymorphism reactions. Multiple regression analysis with adjustments for age and gender were performed and best-fitting models of inheritance were determined.RESULTS: After adjusting for age and sex the TCF7L2 rs7903146 TT genotype was associated with gastric cancer risk under the recessive genetic model(OR = 3.11, 95%CI: 1.22-7.92, P = 0.017). We further investigated the distribution of rs12255372 and rs7903146 genotypes according gastric cancer stratified by degree of differentiation, and we observed that carriers of rs7903146 T allele(CT + TT vs CC) had a significantly increased risk of moderate/well differentiated gastric cancer(dominant model, OR = 2.55, 95%CI: 1.35-4.80, P = 0.004), whereas the rs7903146 TT genotype was associated with poorly differentiated gastric cancer in the recessive model(OR = 3.65, 95%CI: 1.25-10.62, P = 0.018). We did not find association between rs12255372 SNP and the susceptibility of developing gastric cancer. CONCLUSION: TCF7L2 rs7903146 polymorphism is associated with gastric cancer risk in the Venezuelan population, and could be related to determine the degree of differentiation of tumor cells.展开更多
文摘目的观察艾灸对结肠炎相关性结肠癌(CAC)大鼠的干预作用,从嘌呤受体P2X7R与Wnt/b-catenin信号通路探讨可能的效应机制。方法将SD大鼠随机分为正常组、CAC组、隔药灸组、隔姜灸组。CAC组、隔药灸组、隔姜灸组均采用腹腔注射AOM联合DSS法制备CAC大鼠模型,隔药灸与隔姜灸组均取天枢(双)、气海穴进行治疗。记录各组大鼠体质量、疾病活动指数(DAI)和成瘤率;通过HE染色观察艾灸对CAC大鼠结肠损伤的干预效应;通过RT-q PCR和Western Blot技术,检测艾灸对CAC大鼠结肠组织C-myc、Wnt1、b-catenin、GSK-3bm RNA和P2X7R蛋白表达的调节作用。结果与正常组相比,CAC组大鼠体质量显著降低、DAI增高、成瘤率明显增加(P<0.05),结肠组织可见腺管共壁背靠背和筛样结构,高级别腺癌形成。CAC组大鼠结肠组织P2X7R蛋白表达显著下调(P<0.05),C-myc、b-catenin、GSK-3b、Wnt1 m RNA的表达均显著上调(P<0.05)。与CAC组相比,隔药灸组和隔姜灸组大鼠体质量增加,DAI降低(P<0.05),结肠组织P2X7R蛋白表达显著上调,C-myc m RNA下调(P<0.05);隔姜灸组成瘤率明显降低(P<0.05),且Wnt1、b-catenin、GSK-3bm RNA表达显著下调(P<0.05)。结论隔药灸、隔姜灸均能调节CAC大鼠结肠组织P2X7R及C-myc的异常表达,且隔姜灸还能下调CAC大鼠结肠组织Wnt1、b-catenin、GSK-3bm RNA的表达。
基金We thank the Experimental Poultry Farm of the Poultry Institute,Chinese Academy of Agricultural Sciences,for providing experimental materialsThis work was supported by the Key Research and Development Program of China(2017YFE0108000)+1 种基金the National Natural Science Foundation of China(31872341,31572390)the High-Level Talent Support Program of Yangzhou University,China.
文摘Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells. The mechanisms underlying expression and regulation of Lin28A have been well documented, but such information for Lin28B is limited. In this study, a fragment of the Lin28B promoter was cloned, the pEGFP-pLin28B vector was constructed. DF-1 chicken fibroblasts were transfected and the expression of green fluorescent protein (GFP) was measured. Furtherly, Lin28B promoter of different lengths fragments was cloned using the chromosome-walking method and the fragments were ligated into the PGL3-Basic vector, and transfected into DF-1 cells. Results of dual-luciferase reporter assay showed that the core of the Lin28B promoter was included in the sequence from –1 431 to –1 034 bp. The binding sites of the transcription factor TCF7L2 was showed within this sequence by bioinformatics analysis. The promoter activity of Lin28B was downregulated (P<0.05) when the TCF7L2 binding site was mutated. Further experiments suggested that Lin28B promoter activity responded to the activation or inhibition of Wnt signaling. Results of chromatin immunoprecipitation and quantitative PCR showed that β-catenin-TCF7L2 may be enriched in the Lin28B promoter core area. In vivo and in vitro activation or inhibition of Wnt signaling significantly up- or down-regulated (P<0.05) Lin28B expression. H3K4me2 enriched in the promoter of Lin28B, which affected the regulation of Wnt signaling to Lin28B. In conclusion, our results showed that H3K4me2 and Wnt5a/β-catenin/TCF7L2 were the positive regulators of Lin28B expression. Findings of this study may lay a theoretical foundation for illuminating the mechanism underlying Lin28B expression.
文摘AIM: To explore the association between TCF7L2 rs12255372 and rs7903146 single nucleotide polymorphisms(SNPs) and gastric cancer risk in Venezuelan patients.METHODS: We performed a case-control study including 122 paraffin-embedded archived intestinaltype gastric cancer samples and 129 biopsies obtained by superior endoscopy from chronic gastritis patients. Gastric cancer samples were classified according the degree of carcinoma differentiation. Genomic DNA was extracted from tissues, and the two SNPs of TCF7L2 gene(rs12255372 and rs7903146) were genotyped by polymerase chain reaction-restriction fragment length polymorphism reactions. Multiple regression analysis with adjustments for age and gender were performed and best-fitting models of inheritance were determined.RESULTS: After adjusting for age and sex the TCF7L2 rs7903146 TT genotype was associated with gastric cancer risk under the recessive genetic model(OR = 3.11, 95%CI: 1.22-7.92, P = 0.017). We further investigated the distribution of rs12255372 and rs7903146 genotypes according gastric cancer stratified by degree of differentiation, and we observed that carriers of rs7903146 T allele(CT + TT vs CC) had a significantly increased risk of moderate/well differentiated gastric cancer(dominant model, OR = 2.55, 95%CI: 1.35-4.80, P = 0.004), whereas the rs7903146 TT genotype was associated with poorly differentiated gastric cancer in the recessive model(OR = 3.65, 95%CI: 1.25-10.62, P = 0.018). We did not find association between rs12255372 SNP and the susceptibility of developing gastric cancer. CONCLUSION: TCF7L2 rs7903146 polymorphism is associated with gastric cancer risk in the Venezuelan population, and could be related to determine the degree of differentiation of tumor cells.
