Hepatocyte nuclear factor 1 alpha(HNF1A),hepatocyte nuclear factor 4 alpha(HNF4A),and forkhead box protein A2(FOXA2)are key transcription factors that regulate a complex gene network in the liver,cre-ating a regulator...Hepatocyte nuclear factor 1 alpha(HNF1A),hepatocyte nuclear factor 4 alpha(HNF4A),and forkhead box protein A2(FOXA2)are key transcription factors that regulate a complex gene network in the liver,cre-ating a regulatory transcriptional loop.The Encode and ChIP-Atlas databases identify the recognition sites of these transcription factors in many glycosyltransferase genes.Our in silico analysis of HNF1A,HNF4A.and FOXA2 binding to the ten candidate glyco-genes studied in this work confirms a significant enrich-ment of these transcription factors specifically in the liver.Our previous studies identified HNF1A as a master regulator of fucosylation,glycan branching,and galactosylation of plasma glycoproteins.Here,we aimed to functionally validate the role of the three transcription factors on downstream glyco-gene transcriptional expression and the possible effect on glycan phenotype.We used the state-of-the-art clus-tered regularly interspaced short palindromic repeats/dead Cas9(CRISPR/dCas9)molecular tool for the downregulation of the HNF1A,HNF4A,and FOXA2 genes in HepG2 cells-a human liver cancer cell line.The results show that the downregulation of all three genes individually and in pairs affects the transcrip-tional activity of many glyco-genes,although downregulation of glyco-genes was not always followed by an unambiguous change in the corresponding glycan structures.The effect is better seen as an overall change in the total HepG2 N-glycome,primarily due to the extension of biantennary glycans.We propose an alternative way to evaluate the N-glycome composition via estimating the overall complexity of the glycome by quantifying the number of monomers in each glycan structure.We also propose a model showing feedback loops with the mutual activation of HNF1A-FOXA2 and HNF4A-FOXA2 affecting glyco-genes and protein glycosylation in HepG2 cells.展开更多
BACKGROUND A series of long non-coding RNAs(lncRNAs)have been reported to play a crucial role in cancer biology.Some previous studies report that lncRNA CDKN2B-AS1 is involved in some human malignancies.However,its ro...BACKGROUND A series of long non-coding RNAs(lncRNAs)have been reported to play a crucial role in cancer biology.Some previous studies report that lncRNA CDKN2B-AS1 is involved in some human malignancies.However,its role in hepatocellular carcinoma(HCC)has not been fully deciphered.AIM To decipher the role of CDKN2B-AS1 in the progression of HCC.METHODS CDKN2B-AS1 expression in HCC was detected by quantitative real-time polymerase chain reaction.The malignant phenotypes of Li-7 and SNU-182 cells were detected by the CCK-8 method,EdU method,and flow cytometry,respectively.RNA immunoprecipitation was executed to confirm the interaction between CDKN2B-AS1 and E2F transcription factor 1(E2F1).Luciferase reporter assay and chromatin immunoprecipitation were performed to verify the binding of E2F1 to the promoter of G protein subunit alpha Z(GNAZ).E2F1 and GNAZ were detected by western blot in HCC cells.RESULTS In HCC tissues,CDKN2B-AS1 was upregulated.Depletion of CDKN2B-AS1 inhibited the proliferation of HCC cells,and the depletion of CDKN2B-AS1 also induced cell cycle arrest and apoptosis.CDKN2B-AS1 could interact with E2F1.Depletion of CDKN2B-AS1 inhibited the binding of E2F1 to the GNAZ promoter region.Overexpression of E2F1 reversed the biological effects of depletion of CDKN2B-AS1 on the malignant behaviors of HCC cells.CONCLUSION CDKN2B-AS1 recruits E2F1 to facilitate GNAZ transcription to promote HCC progression.展开更多
目的探讨血清壳多糖酶3样蛋白1(chitinase-3-like protein 1,CHI3L1)、甲胎蛋白(alpha fetoprotein,AFP)和γ-谷氨酰转移酶(γ-glutamyl transferase,GGT)检测在乙型肝炎病毒(hepatitis B virus,HBV)感染相关肝癌诊断中的临床应用价值...目的探讨血清壳多糖酶3样蛋白1(chitinase-3-like protein 1,CHI3L1)、甲胎蛋白(alpha fetoprotein,AFP)和γ-谷氨酰转移酶(γ-glutamyl transferase,GGT)检测在乙型肝炎病毒(hepatitis B virus,HBV)感染相关肝癌诊断中的临床应用价值。方法选取2021年7月—2023年7月在玉林市第一人民医院就诊HBV病毒感染相关的50例肝癌患者,50例肝硬化患者,50例慢性乙型肝炎患者以及50名同期健康体检者作为研究对象。比较4组研究对象血清中CHI3L1、AFP、GGT、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)等水平的差异,用受试者工作特征(receiver operating characteristic curve,ROC)曲线评估各指标在肝癌中的诊断价值。结果肝炎组、肝硬化组、肝癌组的CHI3L1、AST、ALT水平均高于对照组,肝癌组AFP、GGT水平高于对照组,差异有统计学意义(P<0.05)。与肝炎组比较,肝硬化组及肝癌组的CHI3L1、AFP、GGT、AST、ALT水平均升高,差异有统计学意义(P<0.05)。与肝硬化组比较,肝癌组的CHI3L1、AFP、GGT、AST水平均升高,差异有统计学意义(P<0.05)。ROC曲线分析显示,CHI3L1、AFP、GGT联合时的曲线下面积(area under the curve,AUC)最大(AUC=0.936)。Spearman相关分析结果显示,CHI3L1与AST呈正相关(r=0.414,P=0.003),AFP与GGT呈正相关(r=0.437,P=0.002),AFP与AST呈正相关(r=0.504,P<0.001),GGT与AST呈正相关(r=0.759,P<0.001),GGT与ALT呈正相关(r=0.636,P<0.001)。结论CHI3L1、AFP及GGT联合检测可提高肝癌的诊断价值,对临床肝癌患者诊疗有重要作用。展开更多
基金the European Structural and Invest-ment Funded Grant"CardioMetabolic"(#KK.01.2.1.02.0321)the Croatian National Centre of Research Excellence in Personalized Healthcare Grant(#KK.01.1.1.01.0010)+2 种基金the European Regional Development Fund Grant,project"CRISPR/Cas9-CasMouse"(#KK.01.1.1.04.0085)the European Structural and Investment Funded Project of Centre of Competence in Molecular Diagnostics(#KK.01.2.2.03.0006)the Croatian National Centre of Research Excellence in Personalized Healthcare Grant(#KK.01.1.1.01.0010).
文摘Hepatocyte nuclear factor 1 alpha(HNF1A),hepatocyte nuclear factor 4 alpha(HNF4A),and forkhead box protein A2(FOXA2)are key transcription factors that regulate a complex gene network in the liver,cre-ating a regulatory transcriptional loop.The Encode and ChIP-Atlas databases identify the recognition sites of these transcription factors in many glycosyltransferase genes.Our in silico analysis of HNF1A,HNF4A.and FOXA2 binding to the ten candidate glyco-genes studied in this work confirms a significant enrich-ment of these transcription factors specifically in the liver.Our previous studies identified HNF1A as a master regulator of fucosylation,glycan branching,and galactosylation of plasma glycoproteins.Here,we aimed to functionally validate the role of the three transcription factors on downstream glyco-gene transcriptional expression and the possible effect on glycan phenotype.We used the state-of-the-art clus-tered regularly interspaced short palindromic repeats/dead Cas9(CRISPR/dCas9)molecular tool for the downregulation of the HNF1A,HNF4A,and FOXA2 genes in HepG2 cells-a human liver cancer cell line.The results show that the downregulation of all three genes individually and in pairs affects the transcrip-tional activity of many glyco-genes,although downregulation of glyco-genes was not always followed by an unambiguous change in the corresponding glycan structures.The effect is better seen as an overall change in the total HepG2 N-glycome,primarily due to the extension of biantennary glycans.We propose an alternative way to evaluate the N-glycome composition via estimating the overall complexity of the glycome by quantifying the number of monomers in each glycan structure.We also propose a model showing feedback loops with the mutual activation of HNF1A-FOXA2 and HNF4A-FOXA2 affecting glyco-genes and protein glycosylation in HepG2 cells.
文摘BACKGROUND A series of long non-coding RNAs(lncRNAs)have been reported to play a crucial role in cancer biology.Some previous studies report that lncRNA CDKN2B-AS1 is involved in some human malignancies.However,its role in hepatocellular carcinoma(HCC)has not been fully deciphered.AIM To decipher the role of CDKN2B-AS1 in the progression of HCC.METHODS CDKN2B-AS1 expression in HCC was detected by quantitative real-time polymerase chain reaction.The malignant phenotypes of Li-7 and SNU-182 cells were detected by the CCK-8 method,EdU method,and flow cytometry,respectively.RNA immunoprecipitation was executed to confirm the interaction between CDKN2B-AS1 and E2F transcription factor 1(E2F1).Luciferase reporter assay and chromatin immunoprecipitation were performed to verify the binding of E2F1 to the promoter of G protein subunit alpha Z(GNAZ).E2F1 and GNAZ were detected by western blot in HCC cells.RESULTS In HCC tissues,CDKN2B-AS1 was upregulated.Depletion of CDKN2B-AS1 inhibited the proliferation of HCC cells,and the depletion of CDKN2B-AS1 also induced cell cycle arrest and apoptosis.CDKN2B-AS1 could interact with E2F1.Depletion of CDKN2B-AS1 inhibited the binding of E2F1 to the GNAZ promoter region.Overexpression of E2F1 reversed the biological effects of depletion of CDKN2B-AS1 on the malignant behaviors of HCC cells.CONCLUSION CDKN2B-AS1 recruits E2F1 to facilitate GNAZ transcription to promote HCC progression.
文摘目的探讨血清壳多糖酶3样蛋白1(chitinase-3-like protein 1,CHI3L1)、甲胎蛋白(alpha fetoprotein,AFP)和γ-谷氨酰转移酶(γ-glutamyl transferase,GGT)检测在乙型肝炎病毒(hepatitis B virus,HBV)感染相关肝癌诊断中的临床应用价值。方法选取2021年7月—2023年7月在玉林市第一人民医院就诊HBV病毒感染相关的50例肝癌患者,50例肝硬化患者,50例慢性乙型肝炎患者以及50名同期健康体检者作为研究对象。比较4组研究对象血清中CHI3L1、AFP、GGT、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)等水平的差异,用受试者工作特征(receiver operating characteristic curve,ROC)曲线评估各指标在肝癌中的诊断价值。结果肝炎组、肝硬化组、肝癌组的CHI3L1、AST、ALT水平均高于对照组,肝癌组AFP、GGT水平高于对照组,差异有统计学意义(P<0.05)。与肝炎组比较,肝硬化组及肝癌组的CHI3L1、AFP、GGT、AST、ALT水平均升高,差异有统计学意义(P<0.05)。与肝硬化组比较,肝癌组的CHI3L1、AFP、GGT、AST水平均升高,差异有统计学意义(P<0.05)。ROC曲线分析显示,CHI3L1、AFP、GGT联合时的曲线下面积(area under the curve,AUC)最大(AUC=0.936)。Spearman相关分析结果显示,CHI3L1与AST呈正相关(r=0.414,P=0.003),AFP与GGT呈正相关(r=0.437,P=0.002),AFP与AST呈正相关(r=0.504,P<0.001),GGT与AST呈正相关(r=0.759,P<0.001),GGT与ALT呈正相关(r=0.636,P<0.001)。结论CHI3L1、AFP及GGT联合检测可提高肝癌的诊断价值,对临床肝癌患者诊疗有重要作用。
