UTX(ubiquitously transcribed TPR gene on the X chromosome)作为H3K27me2/3的去甲基酶,能够调控动物发育过程,并有着重要的生物学作用。根据猪Utx预测序列,扩增Utx的CDS区全长序列。利用Utx的CDS区序列信息,构建pCD513B-CMV-Utx过表...UTX(ubiquitously transcribed TPR gene on the X chromosome)作为H3K27me2/3的去甲基酶,能够调控动物发育过程,并有着重要的生物学作用。根据猪Utx预测序列,扩增Utx的CDS区全长序列。利用Utx的CDS区序列信息,构建pCD513B-CMV-Utx过表达载体,设计并合成干扰shRNA和阴性对照寡核苷酸片段。其中干扰shRNA及阴性对照通过退火形成双链DNA,并插入pCD513B-U6慢病毒干扰载体。采用双酶切的方法和DNA测序鉴定重组载体。通过干扰和过表达载体质粒共转293T细胞,利用qRT-PCR筛选出最有效的干扰序列。重组质粒与其他3种慢病毒辅助包装质粒(pRsv-Rev、pGag-Pol、pVSV-G)共转染293T细胞包装慢病毒。结果表明,本试验成功扩增Utx基因的全长序列,且成功构建了Utx的过表达和干扰载体。qRT-PCR筛选出pCD513B-U6-Utx-shRNA-3干扰效率最显著,干扰效率为70%。本试验成功包装出慢病毒Lenti-Utx-shRNA、Lenti-NC和Lenti-Utx,测定的病毒滴度分别为6.1×107 TU/mL、4.8×107 TU/mL和3.9×105 TU/mL。此试验为进一步在猪源细胞中进行Utx基因的研究奠定基础。展开更多
目的:检测UTX(ubiquitously transcribed TPR gene on the X chromosome)在肾透明细胞癌中的表达及其临床意义。方法:收集临床诊断明确肾透明细胞癌患者组织标本45例,提取组织mRNA,制作组织切片,采用实时定量PCR和免疫组织化学分析UTX...目的:检测UTX(ubiquitously transcribed TPR gene on the X chromosome)在肾透明细胞癌中的表达及其临床意义。方法:收集临床诊断明确肾透明细胞癌患者组织标本45例,提取组织mRNA,制作组织切片,采用实时定量PCR和免疫组织化学分析UTX在肾癌组织和癌旁组织表达的差异,并分析其表达改变与临床病理参数之间关系。结果:实时定量PCR结果显示,UTX mRNA在肾癌组织中的表达明显高于癌旁组织[(0.883 2±0.703 8 vs.0.199 7±0.140 0,P<0.05],免疫组织化学结果同样表明肾癌组织UTX表达水平明显高于癌旁组织(P<0.05),UTX在肾癌组织中的蛋白评分高出癌旁组织4倍[12±4 vs.3±3,P<0.05]。UTX在肾癌组织中的表达与性别、年龄、肿瘤大小、TNM分期无明显关系,但与病理分级关系密切(P=0.004)。结论:UTX在肾透明细胞癌中表达增高,并且与肾癌分级相关。展开更多
Ubiquitously transcribed tetratricopeptide repeat on chromosome X(UTX),also known as lysine(K)-specific demethylase 6A(KDM6A),functions as a tumor suppressor gene or oncogene depending on the tumor type and context.Ho...Ubiquitously transcribed tetratricopeptide repeat on chromosome X(UTX),also known as lysine(K)-specific demethylase 6A(KDM6A),functions as a tumor suppressor gene or oncogene depending on the tumor type and context.However,its tumor-suppressive mechanisms remain largely unknown.Here,we investigated the clinical significance and biological effects of UTX expression in pancreatic ductal adenocarcinoma(PDA)and determined the potential mechanisms of its dysregulation.UTX expression and its association with clinicopathologic characteristics of PDA patients were analyzed using immunohistochemistry.UTX mRNA and protein expression and their regulation in PDA cell lines were measured using quantitative polymerase chain reaction and Western blot analyses.The biological functions of UTX in PDA cell growth,migration,and invasion were determined using gain-and loss-of-function assays with both in vitro and in vivo animal models.UTX expression was reduced in human PDA cell lines and specimens.Low UTX expression was associated with poor differentiation and prognosis in PDA.Forced UTX expression inhibited PDA proliferation,migration,and invasion in vitro and PDA growth and metastasis in vivo,whereas knockdown of UTX expression did the opposite.Mechanistically,UTX expression was trans-activated by GATA6 activation.GATA6-mediated PDA progression could be blocked,at least partially,by silencing UTX expression.In conclusion,loss of GATA6-mediated UTX expression was evident in human PDA and restored UTX expression suppressed PDA growth and metastasis.Thus,UTX is a tumor suppressor in PDA and may serve as a prognostic biomarker and therapeutic target.展开更多
In a recent study.1 published in Signal Transduction and Targeted Therapy,Dr.Yu Liu and collaborators report that the differentiation block in UTX-null leukemia cells can be reverted by an LSD1 inhibitor,highlighting ...In a recent study.1 published in Signal Transduction and Targeted Therapy,Dr.Yu Liu and collaborators report that the differentiation block in UTX-null leukemia cells can be reverted by an LSD1 inhibitor,highlighting additional ways of targeting UTX-deficient malignancies.展开更多
文摘UTX(ubiquitously transcribed TPR gene on the X chromosome)作为H3K27me2/3的去甲基酶,能够调控动物发育过程,并有着重要的生物学作用。根据猪Utx预测序列,扩增Utx的CDS区全长序列。利用Utx的CDS区序列信息,构建pCD513B-CMV-Utx过表达载体,设计并合成干扰shRNA和阴性对照寡核苷酸片段。其中干扰shRNA及阴性对照通过退火形成双链DNA,并插入pCD513B-U6慢病毒干扰载体。采用双酶切的方法和DNA测序鉴定重组载体。通过干扰和过表达载体质粒共转293T细胞,利用qRT-PCR筛选出最有效的干扰序列。重组质粒与其他3种慢病毒辅助包装质粒(pRsv-Rev、pGag-Pol、pVSV-G)共转染293T细胞包装慢病毒。结果表明,本试验成功扩增Utx基因的全长序列,且成功构建了Utx的过表达和干扰载体。qRT-PCR筛选出pCD513B-U6-Utx-shRNA-3干扰效率最显著,干扰效率为70%。本试验成功包装出慢病毒Lenti-Utx-shRNA、Lenti-NC和Lenti-Utx,测定的病毒滴度分别为6.1×107 TU/mL、4.8×107 TU/mL和3.9×105 TU/mL。此试验为进一步在猪源细胞中进行Utx基因的研究奠定基础。
文摘目的:检测UTX(ubiquitously transcribed TPR gene on the X chromosome)在肾透明细胞癌中的表达及其临床意义。方法:收集临床诊断明确肾透明细胞癌患者组织标本45例,提取组织mRNA,制作组织切片,采用实时定量PCR和免疫组织化学分析UTX在肾癌组织和癌旁组织表达的差异,并分析其表达改变与临床病理参数之间关系。结果:实时定量PCR结果显示,UTX mRNA在肾癌组织中的表达明显高于癌旁组织[(0.883 2±0.703 8 vs.0.199 7±0.140 0,P<0.05],免疫组织化学结果同样表明肾癌组织UTX表达水平明显高于癌旁组织(P<0.05),UTX在肾癌组织中的蛋白评分高出癌旁组织4倍[12±4 vs.3±3,P<0.05]。UTX在肾癌组织中的表达与性别、年龄、肿瘤大小、TNM分期无明显关系,但与病理分级关系密切(P=0.004)。结论:UTX在肾透明细胞癌中表达增高,并且与肾癌分级相关。
基金supported by the Jiangxi Science Fund for Distinguished Young Scholars(China)(No.20212ACB216012)the Funding Program for Academic and Technical Leaders of Main Subjects in Jiangxi Province,China(No.20213BCJ22009 to H.Q.Zhang)+4 种基金the National Natural Science Foundation of China(No.81460372 to H.Q.Zhang,No.81960528 to S.Zheng)the Hainan Province Science and Technology special fund(China)(ZDYF2020132 to S.Zheng)the Innovation Platform for Academicians of Hainan Province(China)(YSPTZX202208 to S.Zheng)Hainan Province Clinical Medical Center(QWYH2021276)the Cardiovascular Disease Research Science Innovation Group of Hainan Medical University(China).
文摘Ubiquitously transcribed tetratricopeptide repeat on chromosome X(UTX),also known as lysine(K)-specific demethylase 6A(KDM6A),functions as a tumor suppressor gene or oncogene depending on the tumor type and context.However,its tumor-suppressive mechanisms remain largely unknown.Here,we investigated the clinical significance and biological effects of UTX expression in pancreatic ductal adenocarcinoma(PDA)and determined the potential mechanisms of its dysregulation.UTX expression and its association with clinicopathologic characteristics of PDA patients were analyzed using immunohistochemistry.UTX mRNA and protein expression and their regulation in PDA cell lines were measured using quantitative polymerase chain reaction and Western blot analyses.The biological functions of UTX in PDA cell growth,migration,and invasion were determined using gain-and loss-of-function assays with both in vitro and in vivo animal models.UTX expression was reduced in human PDA cell lines and specimens.Low UTX expression was associated with poor differentiation and prognosis in PDA.Forced UTX expression inhibited PDA proliferation,migration,and invasion in vitro and PDA growth and metastasis in vivo,whereas knockdown of UTX expression did the opposite.Mechanistically,UTX expression was trans-activated by GATA6 activation.GATA6-mediated PDA progression could be blocked,at least partially,by silencing UTX expression.In conclusion,loss of GATA6-mediated UTX expression was evident in human PDA and restored UTX expression suppressed PDA growth and metastasis.Thus,UTX is a tumor suppressor in PDA and may serve as a prognostic biomarker and therapeutic target.
文摘In a recent study.1 published in Signal Transduction and Targeted Therapy,Dr.Yu Liu and collaborators report that the differentiation block in UTX-null leukemia cells can be reverted by an LSD1 inhibitor,highlighting additional ways of targeting UTX-deficient malignancies.
