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酵母双杂交系统鉴定hTCF4相互作用 被引量:3
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作者 林桂亭 张志文 +3 位作者 藏桐 辛殿旗 常智杰 郭应禄 《科学通报》 EI CAS CSCD 北大核心 2000年第8期851-856,共6页
利用酵母双杂交试验技术,鉴定了Wnt/Frizzled信号通路中的关键信号分子TCF4自身的相互作用.通过 TCF4突变体的同源相互作用的双杂交反应(two hybridization),确定了转录因子TCF4在发挥作... 利用酵母双杂交试验技术,鉴定了Wnt/Frizzled信号通路中的关键信号分子TCF4自身的相互作用.通过 TCF4突变体的同源相互作用的双杂交反应(two hybridization),确定了转录因子TCF4在发挥作用过程中形成双聚体,推测以HLH或LZ的结构花式(motif)调控下游基因的表达. 展开更多
关键词 Wnt/Frizzled 酵母双杂交 鉴定 htcf4 相互作用
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hTCF_4酵母双杂交载体的构建
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作者 林桂亭 张志文 +2 位作者 藏桐 辛殿旗 郭应禄 《潍坊医学院学报》 2002年第1期18-19,共2页
目的 研究Wnt Frizzled信号传导通路上的主要信号分子T细胞因子 (TCF4)在细胞核内转导的辅激活物和辅阻遏物。方法 通过PCR法以真核表达载体pCDNA TCF4为模板 ,扩增获得了15 0 0bp的TCF4基因的末端及DNA结合域片段 ,经过SalI ,NotI酶... 目的 研究Wnt Frizzled信号传导通路上的主要信号分子T细胞因子 (TCF4)在细胞核内转导的辅激活物和辅阻遏物。方法 通过PCR法以真核表达载体pCDNA TCF4为模板 ,扩增获得了15 0 0bp的TCF4基因的末端及DNA结合域片段 ,经过SalI ,NotI酶切后 ,基因重组的方法定向克隆于酵母双杂交载体pPC86和pDBleu中 ,构建形成pDBleu TCF4和pPC86 TCF4真核表达载体。结果 经转化 ,提取质粒酶切鉴定表明构建成功。结论 hTCF4酵母双杂交载体的构建成功为Wnt Frizzled信号通路的深入研究奠定了基础。 展开更多
关键词 酵母双杂交 载体 亚克隆 htcf4 pDBleu-TCF4 pPC86-TCF4
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Interaction of hTCF4 by yeast two-hybrid system 被引量:1
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作者 LIN Guiting ZHANG Zhiwen +3 位作者 ZANG long XIN Dianqi CHANG Zhijie GUO Yinglu 《Chinese Science Bulletin》 SCIE EI CAS 2000年第21期1973-1977,共5页
To investigate the interaction of hTCF4, the yeast two-hybrid system has been used for testing the interaction of mutants of hTCF4 with themselves. Mutants of hTCF4 (hTCF4 Ⅰ ) and hTCF4 Ⅱ) have been obtained by poly... To investigate the interaction of hTCF4, the yeast two-hybrid system has been used for testing the interaction of mutants of hTCF4 with themselves. Mutants of hTCF4 (hTCF4 Ⅰ ) and hTCF4 Ⅱ) have been obtained by polymerase chain reaction (PCR). Bait (hTCF4 Ⅰ -pDBLeu and hTCF4Ⅱ-pDBLeu) and prey (hTCF4 Ⅰ -pPC86 and hTCF4Ⅱ-pPC86) have been constructed by DNA recombination for yeast two-hybrid. Interaction of hTCF4Ⅱ with itself is found in the reverse yeast two-hybrid system (GIBCOBRL Co.). However, no interactions are found in hTCF4Ⅱ with hTCF4 Ⅰ and in hTCF4 Ⅰ with hTCF4 Ⅰ . These results suggest that hTCF4 could interact with itself to form homodimer or homocopolymer and perform the transcriptional activating function through LZ or HLH motifs in nucleus of renal cell carcinoma. 展开更多
关键词 Wnt/Frizzled htcf4 YEAST two-hybrid.
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miRNA-320a、hTcf-4、β-catenin在肝癌组织的表达及临床意义 被引量:2
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作者 祁蕙燕 郭剑 程燕莉 《分子诊断与治疗杂志》 2020年第10期1401-1405,1410,共6页
目的探讨肝癌组织微小RNA(miRNA-320a)、人T细胞因子4基因(hTcf-4)、β联蛋白(β-catenin)表达及与患者病理特征、预后的关系。方法选取2017年6月至2019年8月本院收治的122例肝癌患者进行回顾性研究,比较miRNA-320a、hTcf-4 mRNA、β-ca... 目的探讨肝癌组织微小RNA(miRNA-320a)、人T细胞因子4基因(hTcf-4)、β联蛋白(β-catenin)表达及与患者病理特征、预后的关系。方法选取2017年6月至2019年8月本院收治的122例肝癌患者进行回顾性研究,比较miRNA-320a、hTcf-4 mRNA、β-catenin在肝癌组织与癌旁组织中表达,并比较各指标阳性与阴性表达患者病理特征,对数据进行统计学分析。结果miRNA-320a表达水平与分化程度呈正相关(r=6.894,P<0.05),与肿瘤T分期、肿瘤大小、淋巴结转移呈负相关(r=-5.771、-6.351、-7.086,P<0.05);hTcf-4 mRNA(r=-0.608,P<0.05)、β-catenin表达水平(r=-0.626,P<0.05)与分化程度呈负相关,与肿瘤T分期(r=0.617、0.449,P<0.05)、肿瘤大小(r=0.544、0.351,P<0.05)、淋巴结转移呈正相关(r=0.589、0.539,P<0.05)。结论肝癌组织miRNA-320a呈低表达,hTcf-4、β-catenin呈高表达,均与分化程度、肿瘤T分期、肿瘤大小、淋巴结转移有关,并能预测患者预后。 展开更多
关键词 肝癌组织 miRNA-320a htcf-4 β联蛋白 病理特征
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Novel mutations and sequence variants in exons 3-9 of human T Cell Factor-4 gene in sporadic rectal cancer patients stratified by microsatellite instability 被引量:1
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作者 Wen-Jian Meng Ling Wang +9 位作者 Chao Tian Yong-Yang Yu Bing Zhou Jun Gu Qing-Jie Xia Xiao-Feng Sun Yuan Li Rong Wang Xue-Lian Zheng Zong-Guang Zhou 《World Journal of Gastroenterology》 SCIE CAS CSCD 2007年第27期3747-3751,共5页
AIM: To establish the role of human T Cell Factor-4 (hTCF-4) gene exons 3-9 mutation status in association with sporadic rectal cancer with microsatellite instability (MSI). METHODS: Microsatellite markers were ... AIM: To establish the role of human T Cell Factor-4 (hTCF-4) gene exons 3-9 mutation status in association with sporadic rectal cancer with microsatellite instability (MSI). METHODS: Microsatellite markers were genotyped in 93 sporadic rectal cancer patients. Eleven cases were found to be high-frequency MSI (MSI-H). Sequence analysis of the coding region of the exons 3-9 of hTCF-4 gene was carried out for the 11 MSI-H cases and 10 controls (5 microsatellite stability (MSS) cases and 5 cases with normal mucosa). The sequencing and MSI identification were used. RESULTS: Several novel mutations and variants were revealed. In exon 4, one is a 4-position continuous alteration which caused amino acid change from Q131T and S132I (391insA, 392 G 〉 A, 393 A 〉 G and 395delC) and another nucleotide deletion (395delC) is present in MSI-H cases (5/10 and 4/10, respectively) but completely absent in the controls.CONCLUSION: Novel mutations in exon 4 of hTCF-4 gene were revealed in this study, which might be of importance in the pathogenesis of sporadic rectal cancer patients with MSI-H. 展开更多
关键词 htcf-4 Sporadic rectal cancer Microsatellite instability Mutation analysis
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肝细胞肝癌中T细胞转录因子4基因表达及突变 被引量:1
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作者 江颖 周信达 +1 位作者 刘银坤 吴欣 《中国癌症杂志》 CAS CSCD 2002年第1期5-8,共4页
目的 :研究人T细胞转录因子 4(hTcf 4 )基因在肝细胞肝癌中表达及突变和肝癌发生发展的关系。方法 :用逆转录 -聚合酶链反应 (RT PCR)检测了 32例肝癌组织及癌旁组织中hTcf 4基因表达 ,并用聚合酶链反应 -单链构像多态性分析 (PCR SSCP... 目的 :研究人T细胞转录因子 4(hTcf 4 )基因在肝细胞肝癌中表达及突变和肝癌发生发展的关系。方法 :用逆转录 -聚合酶链反应 (RT PCR)检测了 32例肝癌组织及癌旁组织中hTcf 4基因表达 ,并用聚合酶链反应 -单链构像多态性分析 (PCR SSCP)方法检测了肝癌组织中hTcf 4外显子 1、4、9、15的突变情况。结果 :hTcf 4mRNA在癌组织及癌旁组织中表达率分别为 90 .6 %和 71.9%。且癌组织中表达水平明显高于癌旁组织 [(0 .71±0 .13)vs (0 .2 9± 0 .0 5 ) ,P <0 .0 0 1]。同时 ,此基因在包膜不完整及有转移的癌组织表达增高更为显著。 32例肝癌中仅检测到 2例hTcf 4基因外显子 15突变 ,突变率为 6 .2 5 %。结论 :hTcf 4基因在肝细胞肝癌 ,尤其伴转移肝癌组织中表达增高 ,并与肝癌侵袭转移有关 ,而该基因突变与肝癌关系不密切。 展开更多
关键词 肝肿瘤 T淋巴细胞 逆转录聚合酶链反应 基因突变 htcf-4 肝细胞癌
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Association of T cell factor-4 mRNA expression with clinicopathological characteristics of colorectal cancer
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作者 Junfeng Wang Zongguang Zhou Dianchao Wu 《The Chinese-German Journal of Clinical Oncology》 CAS 2008年第10期584-586,共3页
Objective: To study the expression of T-cell transcription factor-4 (Tcf-4) gene in human colorectal cancer (CRC) and the relationship between Tcf-4 gene and progression of CRC. Methods: Quantitative real time R... Objective: To study the expression of T-cell transcription factor-4 (Tcf-4) gene in human colorectal cancer (CRC) and the relationship between Tcf-4 gene and progression of CRC. Methods: Quantitative real time RT-PCR was used to de- tect the expression of Tcf-4 mRNA in 31 CRC and para-cancerous tissues. The correlation of expression of the Tcf-4 gene with clinicopathologicel characteristics of CRC was also analyzed. Results: Compared with normal tissues, up-regulation of Tcf4 mRNAwas observed in 31 CRC tissues (t = 2.188, P = 0.037). The expression values of Tcf-4 mRNA in Dukes A, B, C and D were 8.38 + 1.92, 6.93 + 1.44, 6.00 + 1.26 and 3.50 + 0.71 respectively. Analyzed by one-way ANOVA, there were significant differences in the expressions of Tcf-4 in different Dukes stages (F = 6.454, P = 0.002). Moreover, the over-expression of Tcf-4 was significantly correlated with lymph node metastases. However the expression of Tcf-4 was no correlation with the age, gender of patients and cell differentiation of CRC in this study. Conclusion: The higher expression level of Tcf-4 mRNA in CRC tissues suggested that the over-expression of Tcf-4 gene may be related to the development and invasion of CRC. 展开更多
关键词 colorectal cancer Tcf-4 mRNA quantitative real time RT-PCR
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结直肠癌组织中hTcf-4 mRNA表达的研究
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作者 王俊锋 周总光 +1 位作者 郑雪莲 吴殿超 《华西医学》 CAS 2005年第2期278-279,共2页
目的:研究hTcf-4mRNA在结直肠癌组织及癌旁正常组织的表达,探讨其在结直肠癌发生发展中的作用。方法:用荧光定量PCR检测31例结直肠癌标本及相应的癌旁正常组织hTcf-4mRNA的表达。结果:31例癌组织hTcf-4mRNA表达高于癌旁正常组织(P<0.... 目的:研究hTcf-4mRNA在结直肠癌组织及癌旁正常组织的表达,探讨其在结直肠癌发生发展中的作用。方法:用荧光定量PCR检测31例结直肠癌标本及相应的癌旁正常组织hTcf-4mRNA的表达。结果:31例癌组织hTcf-4mRNA表达高于癌旁正常组织(P<0.05);Dukes不同分期结直肠癌hTcf-4mRNA表达随着浸润深度的加深其表达有增高趋势(P<0.05)。结论:hTcf-4mRNA在结直肠癌组织中表达增高,并与结直肠癌侵袭转移有关,可作为判定结直肠癌转移及预后的参考指标。 展开更多
关键词 hTef-4 结直肠癌 荧光定量PCR MRNA表达 肿瘤 癌组织
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Association of hTcf-4 gene expression and mutation with clinicopathological characteristics of hepatocellular carcinoma 被引量:23
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作者 Ying Jiang Xin-Da Zhou Yin-Kun Liu Xin Wu Xiao-Wu Huang Liver Cancer Institute,Zhong Shah Hospital,Fudan University,Shanghai 200032,China 《World Journal of Gastroenterology》 SCIE CAS CSCD 2002年第5期804-807,共4页
AIM: Hepatocellular carcinoma(HCC) is a significant healthproblem in China. But the molecular mechanisms of HCCremains unclear. APC/β-Catenin/Tcf signaling pathway, alsoknown as Wnt pathway, plays a critical role in ... AIM: Hepatocellular carcinoma(HCC) is a significant healthproblem in China. But the molecular mechanisms of HCCremains unclear. APC/β-Catenin/Tcf signaling pathway, alsoknown as Wnt pathway, plays a critical role in thedevelopment and oncogenesis. As little is known about thealteration of human T-cell transcription factor-4 (hTcf-4) genein HCC, it is of interest to study the expression and mutationof hTcf-4 gene in HCC and the relationship between hTcf-4gene and progression of HCC.METHODS: Reverse transcription-polymerase chain reaction(RT-PCR) method was used to detect the expression of hTcf-4 mRNA in 32 HCC and para-cancerous tissues and 5 normalliver tissues. PCR-single strand conformation polymorphism(PCR-SSCP) method was used to detect the mutation ofhTcf-4 exons 1, 4, 9 and 15 in HCC. The correlation ofexpression and mutation of the hTcf-4 gene withclinicopathological characteristics of HCC was also analyzed.RESULTS: RT-PCR showed that the expression rate of hTcf-4 mRNA in HCC, para-cancerous tissues and normal livertissues was 90.6 %, 71.9 % and 80 %, respectively. Thegene expression level in tumor was 0.71±0.13, much higherthan that in para-cancerous liver 0.29±0.05 and normal liver0.26±0.05 (P<0.001), although there was no significantdifference in gene expression level between para-canceroustissues and normal liver (P>0.05). Furthermore, hTcf-4 geneexpression was closely associated with tumor capsule statusand intrahepatic metastasis of HCC. On SSCP, 2 of 32 casesof HCC (6.25 %) displayed characteristic mutational mobilityshifts in exon 15 of the hTcf-4 gene. No abnormal shiftingbands were observed in para-cancerous tissues.CONCLUSION: The high expression level of hTcf-4 in HCC,especially in tumors with metastasis, suggests that the over-expression of hTcf-4 gene may be closely associated withdevelopment and progression of HCC, but the mutation ofthis gene seemed to play less important role in this respect. 展开更多
关键词 肝癌 临床病理学 基因突变 RT-PCR htcf-4基因
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hTcf-4基因在肝癌细胞系中的表达及其突变的研究 被引量:1
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作者 江颖 周信达 +4 位作者 刘银坤 吴欣 孙瑞霞 赵燕 陈洁 《中华实验外科杂志》 CAS CSCD 北大核心 2002年第2期107-109,共3页
目的 研究hTcf 4基因在肝癌细胞系中的表达及突变的情况 ,以探讨hTcf 4基因与肝癌发生及侵袭转移的关系。方法 用逆转录 聚合酶链反应 (RT PCR)检测 5种肝癌细胞系和肺癌细胞系、乳腺癌细胞系、胃癌细胞系及正常肝细胞中hTcf 4基因表... 目的 研究hTcf 4基因在肝癌细胞系中的表达及突变的情况 ,以探讨hTcf 4基因与肝癌发生及侵袭转移的关系。方法 用逆转录 聚合酶链反应 (RT PCR)检测 5种肝癌细胞系和肺癌细胞系、乳腺癌细胞系、胃癌细胞系及正常肝细胞中hTcf 4基因表达 ,并用聚合酶链反应 单链构像多态性分析 (PCR SSCP)方法检测肝癌细胞系中hTcf 4外显子 1、4、9、1 5的突变情况。结果 hTcf 4mRNA在不同细胞系中均有表达。但不同细胞系的表达水平差异有显著性 (P <0 .0 5)。低转移肝癌细胞系HepG2、Hep3B、SMMC 772 1、BEL 740 2的表达水平分别为0 .70 5± 0 .0 50 ,0 .745± 0 .0 65 ,0 .71 0± 0 .0 78,0 .746± 0 .0 84,均高于正常肝细胞 0 .452± 0 .0 38(P <0 .0 0 1 )及肺癌细胞系 0 .531± 0 .0 83、乳腺癌细胞系 0 .578± 0 .0 64、胃癌细胞系 0 .50 3± 0 .0 33等非肝癌细胞系 (P <0 .0 5)。而高转移肝癌细胞系MHCC97表达水平为 0 .92 4± 0 .0 74,又高于低转移肝癌细胞系(P <0 .0 1 )。 5种肝癌细胞系中 ,只有MHCC 97hTcf 4基因第 1 5外显子处存在点突变。结论 hTcf 展开更多
关键词 T淋巴细胞 肝细胞癌 基因表达 基因突变 htcf-4基因
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