Background:Hyperuricemia is a metabolic disorder which is characterized by increased serum uric acid levels,which can contribute to serious health issues such as gout,cardiovascular disease,and kidney damage.Epigeneti...Background:Hyperuricemia is a metabolic disorder which is characterized by increased serum uric acid levels,which can contribute to serious health issues such as gout,cardiovascular disease,and kidney damage.Epigenetic modifications,for example,DNA methylation,exert a crucial function in gene regulation and have been implicated in various metabolic disorders.The ATP-Binding Cassette Subfamily G Member 2(ABCG2)gene is involved in uric acid excretion,and its expression can be influenced by methylation of its promoter region.Methods:This study involved the design of three guide RNA(gRNA)sequences targeting specific CpG sites within the ABCG2 promoter region.Using the Clustered Regularly Interspaced Short Palindromic Repeats/dead Cas9-Ten-Eleven Translocation 1(CRISPR/dCas9-TET1)system,these gRNAs were employed to guide targeted demethylation of the ABCG2 promoter in cell models.A non-targeting gRNA served as a negative control.The methylation status of the ABCG2 promoter and its effect on gene expression were assessed using bisulfite sequencing and qRT-PCR.Results:Among the gRNAs tested,gRNA2 and gRNA3 effectively guided the dCas9-TET1 complex to the ABCG2 promoter,resulting in significant demethylation.gRNA2 showed the most pronounced effect,leading to a substantial increase in ABCG2 expression.Clinical data analysis revealed that adolescents with hyperuricemia had higher uric acid levels compared to healthy controls,and a higher proportion of the hyperuricemia group reported a high-protein diet,suggesting a link between diet and ABCG2 methylation.Conclusion:The findings demonstrate that targeted demethylation of the ABCG2 promoter can significantly upregulate its expression,which may help modulate uric acid levels.These results indicate that dietary factors,such as a high-protein diet,could influence ABCG2 methylation and thus impact hyperuricemia.Advanced research is necessary to explore the therapeutic potential of aiming at epigenetic modifications for the treatment of hyperuricemia.展开更多
背景与目的:三磷酸腺苷结合盒转运体成员ABCG2(ATP-binding cassette superfamily G member2)是源于造血干细胞的标志物之一,其在神经胶质瘤发生发展相关组织和细胞中的表达情况还不清楚。本研究检测ABCG2在不同恶性程度人脑胶质瘤组织...背景与目的:三磷酸腺苷结合盒转运体成员ABCG2(ATP-binding cassette superfamily G member2)是源于造血干细胞的标志物之一,其在神经胶质瘤发生发展相关组织和细胞中的表达情况还不清楚。本研究检测ABCG2在不同恶性程度人脑胶质瘤组织标本、裸小鼠移植瘤标本、体外细胞系球体和胶质瘤干细胞球体中的表达情况并分析其意义。方法:制作布有不同恶性程度人脑胶质瘤组织标本、裸小鼠移植瘤标本、体外细胞系球体和胶质瘤干细胞球体等的组织芯片,用免疫组化方法检测ABCG2在组织芯片中的表达情况。结果:在71例人脑胶质瘤组织标本中ABCG2的阳性率为26.8%,其中Ⅰ级11.1%,Ⅱ级8.0%,Ⅲ级43.5%,Ⅳ级42.9%;Ⅰ～Ⅱ级与Ⅲ～Ⅳ级相比差异具有统计学意义(%2=10.710,P=0.001)。在神经干细胞、裸小鼠移植瘤、胶质瘤干细胞球体表达率为100%。在多种正常组织中亦有不同程度的表达。在胶质瘤临床标本中ABCG2阳性细胞呈亲血管分布。结论:ABCG2在胶质瘤干细胞、恶性程度高的胶质瘤组织标本和移植瘤组织中高表达,并且呈亲血管分布。展开更多
目的:明确结直肠癌组织中ABCG2(ATP-binding cassette superfamily G member 2,ABCG2)的表达及其与患者临床病理特征、以伊立替康(CPT-11)为基础化疗疗效的相关性,从而为结直肠癌患者实现个体化治疗提供理论依据。方法:筛选1996年1月至2...目的:明确结直肠癌组织中ABCG2(ATP-binding cassette superfamily G member 2,ABCG2)的表达及其与患者临床病理特征、以伊立替康(CPT-11)为基础化疗疗效的相关性,从而为结直肠癌患者实现个体化治疗提供理论依据。方法:筛选1996年1月至2011年12月于北京大学肿瘤医院行CPT-11化疗的晚期结直肠癌患者,收集患者完整病史资料及肿瘤组织样本。用免疫组织化学的方法检测样本中ABCG2蛋白表达情况。分析ABCG2蛋白表达与患者临床病理特征、CPT-11化疗疗效的相关性。结果:1)免疫组织化学染色显示ABCG2在结直肠癌组织中表达阳性率达93.2%,癌旁正常肠黏膜ABCG2蛋白阳性表达率为43.6%,癌组织较癌旁正常黏膜组织的表达水平明显增高(P<0.05)。ABCG2表达与患者年龄、性别、病理分化程度、病变部位、肝转移均无明显相关性。2)ABCG2蛋白表达与CPT-11化疗疗效无显著相关(P>0.05)。结论:在应用伊立替康为基础方案化疗的结直肠癌患者中,癌组织中ABCG2蛋白表达水平与患者化疗疗效无明显相关。展开更多
基金supported by the Science and Technology Plan Program of Yantian District of Shenzhen(YTWS20200208)Project funding department is Shenzhen Yantian District Science and Technology Innovation Bureau。
文摘Background:Hyperuricemia is a metabolic disorder which is characterized by increased serum uric acid levels,which can contribute to serious health issues such as gout,cardiovascular disease,and kidney damage.Epigenetic modifications,for example,DNA methylation,exert a crucial function in gene regulation and have been implicated in various metabolic disorders.The ATP-Binding Cassette Subfamily G Member 2(ABCG2)gene is involved in uric acid excretion,and its expression can be influenced by methylation of its promoter region.Methods:This study involved the design of three guide RNA(gRNA)sequences targeting specific CpG sites within the ABCG2 promoter region.Using the Clustered Regularly Interspaced Short Palindromic Repeats/dead Cas9-Ten-Eleven Translocation 1(CRISPR/dCas9-TET1)system,these gRNAs were employed to guide targeted demethylation of the ABCG2 promoter in cell models.A non-targeting gRNA served as a negative control.The methylation status of the ABCG2 promoter and its effect on gene expression were assessed using bisulfite sequencing and qRT-PCR.Results:Among the gRNAs tested,gRNA2 and gRNA3 effectively guided the dCas9-TET1 complex to the ABCG2 promoter,resulting in significant demethylation.gRNA2 showed the most pronounced effect,leading to a substantial increase in ABCG2 expression.Clinical data analysis revealed that adolescents with hyperuricemia had higher uric acid levels compared to healthy controls,and a higher proportion of the hyperuricemia group reported a high-protein diet,suggesting a link between diet and ABCG2 methylation.Conclusion:The findings demonstrate that targeted demethylation of the ABCG2 promoter can significantly upregulate its expression,which may help modulate uric acid levels.These results indicate that dietary factors,such as a high-protein diet,could influence ABCG2 methylation and thus impact hyperuricemia.Advanced research is necessary to explore the therapeutic potential of aiming at epigenetic modifications for the treatment of hyperuricemia.
文摘背景与目的:三磷酸腺苷结合盒转运体成员ABCG2(ATP-binding cassette superfamily G member2)是源于造血干细胞的标志物之一,其在神经胶质瘤发生发展相关组织和细胞中的表达情况还不清楚。本研究检测ABCG2在不同恶性程度人脑胶质瘤组织标本、裸小鼠移植瘤标本、体外细胞系球体和胶质瘤干细胞球体中的表达情况并分析其意义。方法:制作布有不同恶性程度人脑胶质瘤组织标本、裸小鼠移植瘤标本、体外细胞系球体和胶质瘤干细胞球体等的组织芯片,用免疫组化方法检测ABCG2在组织芯片中的表达情况。结果:在71例人脑胶质瘤组织标本中ABCG2的阳性率为26.8%,其中Ⅰ级11.1%,Ⅱ级8.0%,Ⅲ级43.5%,Ⅳ级42.9%;Ⅰ～Ⅱ级与Ⅲ～Ⅳ级相比差异具有统计学意义(%2=10.710,P=0.001)。在神经干细胞、裸小鼠移植瘤、胶质瘤干细胞球体表达率为100%。在多种正常组织中亦有不同程度的表达。在胶质瘤临床标本中ABCG2阳性细胞呈亲血管分布。结论:ABCG2在胶质瘤干细胞、恶性程度高的胶质瘤组织标本和移植瘤组织中高表达,并且呈亲血管分布。
文摘目的:明确结直肠癌组织中ABCG2(ATP-binding cassette superfamily G member 2,ABCG2)的表达及其与患者临床病理特征、以伊立替康(CPT-11)为基础化疗疗效的相关性,从而为结直肠癌患者实现个体化治疗提供理论依据。方法:筛选1996年1月至2011年12月于北京大学肿瘤医院行CPT-11化疗的晚期结直肠癌患者,收集患者完整病史资料及肿瘤组织样本。用免疫组织化学的方法检测样本中ABCG2蛋白表达情况。分析ABCG2蛋白表达与患者临床病理特征、CPT-11化疗疗效的相关性。结果:1)免疫组织化学染色显示ABCG2在结直肠癌组织中表达阳性率达93.2%,癌旁正常肠黏膜ABCG2蛋白阳性表达率为43.6%,癌组织较癌旁正常黏膜组织的表达水平明显增高(P<0.05)。ABCG2表达与患者年龄、性别、病理分化程度、病变部位、肝转移均无明显相关性。2)ABCG2蛋白表达与CPT-11化疗疗效无显著相关(P>0.05)。结论:在应用伊立替康为基础方案化疗的结直肠癌患者中,癌组织中ABCG2蛋白表达水平与患者化疗疗效无明显相关。
