TG-interacting factors （TGIFs） belong to a family of TALE-homeodomain proteins including TGIF, TGIF2, and TGIF2LX/Y （TGIF2 like on X or Y chromosome） in human. They potentially play important functions in various ...TG-interacting factors （TGIFs） belong to a family of TALE-homeodomain proteins including TGIF, TGIF2, and TGIF2LX/Y （TGIF2 like on X or Y chromosome） in human. They potentially play important functions in various tissues during development. Mutations in TGIF are frequently associated with malformation of forebrain and facial structures; TGIF2 proteins are over-expressed in many ovarian cancer cell lines; and TGIF2LX/Y are specifically expressed in adult testis. The molecular functions of these proteins have been investigated mostly in cultured cells. TGIF and TGIF2 have been found as transcriptional repressors that modulate TGF-beta signaling. However these findings are far from sufficient to explain their mutant phenotypes or expression patterns, and the functions of TGIF2LX/Y have never been reported. Here we use Drosophila as a model system to explore the functions of TGIF family proteins in vivo. We observed in fly tissues such as fat body, epithelia, and neuronal cells, that expressing human TGIF2 or human TGIF2LX generally inhibited cell growth in size and number. Co-expressing Drosophila Myc, Cyclin E, or human c-MycS partially rescued the growth inhibition induced by human TGIFs, whereas activated insulin pathway signaling did not. Taken together, we provide in vivo evidence for the potential functions of human TGIF2 and TGIF2LX in growth control. Additionally, we confirmed that Drosophila TGIFs are transcriptional activators by assaying their activities in spermatogenesis.展开更多
Excess ammonia(NH_(3))in the circulation of dairy animals can reduce animal health and the quality of products for human consumption.To develop effective prevention and treatment methods,it is essential to examine the...Excess ammonia(NH_(3))in the circulation of dairy animals can reduce animal health and the quality of products for human consumption.To develop effective prevention and treatment methods,it is essential to examine the molecular mechanisms through which excess NH_(3) may affect the mammary gland.The present study used bovine mammary epithelial cells(BMECs)to evaluate the effects of exogenous NH_(4)Cl on the abundance of circular RNAs(circRNAs)using high-throughput sequencing.Among the identified circRNAs,circ02771 was the most significantly upregulated by exogenous NH_(4)Cl(P<0.05),with a fold change of 4.12.The results of the apoptosis and proliferation assays,transmission electron microscopy,H&E staining,and immunohistochemistry revealed that circ02771 increased apoptosis and inflammation.A double luciferase reporter assay revealed that circ02771 targeted miR-194b,and the overexpression of circ02771(pcDNA-circ02771)reduced(P<0.05)the expression of miR-194b and led to apoptosis and inflammation.Circ02771 also enhanced the expression of transforming growth factor beta-induced factor homeobox 1(TGIF1),which is a target gene of miR-194b.Overall,this study suggests that the circ02771/miR-194b/TGIF1 axis plays a role in mediating the effects of NH_(4)Cl on BMECs.Therefore,this axis provides a novel target to help control hazards within the mammary gland from high circulating NH_(4)Cl levels.展开更多
Endometritis affects the reproductive capacity of dairy cows and leads to serious economic losses in dairy farming.Clarification of the pathogenesis of endometritis is necessary to improve the reproductive efficiency ...Endometritis affects the reproductive capacity of dairy cows and leads to serious economic losses in dairy farming.Clarification of the pathogenesis of endometritis is necessary to improve the reproductive efficiency of dairy cows.Exosomes and their miRNAs have been proven to play an important role in inflammatory regulation.Exosomal miR-218 is a differentially expressed miRNA found in endometrial epithelial cells(EECs)under endometrial inflammation.Therefore,we investigated the expression of miR-218 in the uterine tissue of dairy cows,lipopolysaccharide(LPS)treated EECs,exosomal vesicles,and regulation of exosomal miR-218 by targeting TGIF-2 inducible factor homology frame 2(TGIF2)/transforming growth factor-beta(TGF-β).The expression of miR-218 was suppressed in inflammatory uterine tissues and LPS treated EECs.The expression of TGIF2 and TGF-βin inflammatory uterine tissues and LPS treated EECs was significantly higher than those in healthy uterine tissues and EECs(p<0.01).Interestingly,miR-218 derived from donor cells was found to regulate the expression of the target gene TGIF2 in recipient cells through the fusion of exosomes.Concurrently,the expression of its target gene TGIF2 was also suppressed by miR-218 in donor cells resulting in fewer TGIF2 being transported into recipient cells with exosomal fusion.This may be a novel mechanism of miRNAs-mediated regulation and provides a new reference for analyzing the pathogenesis of endometritis in dairy cows.展开更多
Objective: To observe the effect of the expression of transforming growth factor β-homologous inducible factor 1 (TGIF1) on the expression of epithelial cadherin (E-cadherin) and human Twist-related protein 1 (Twist1...Objective: To observe the effect of the expression of transforming growth factor β-homologous inducible factor 1 (TGIF1) on the expression of epithelial cadherin (E-cadherin) and human Twist-related protein 1 (Twist1) in breast cancer cells. Methods: Total of twenty-four 6-week-old female SPF Balb/c mice were randomly divided into a control group, a TGIF1-silencing group, a TGIF1-normal group, and a TGIF1-overexpression group. In the TGIF1-silencing group, 4T1 breast cancer cells were interfered by lentivirus shRNA (H) lentiviral particles (sc-36659-v) to construct a breast cancer model. TGIF1-normal group used breast cancer cells (4T1) to construct a mouse model of breast cancer. And the TGIF1-overexpression group used 4T1 breast cancer cells with TGIF1 overexpression to establish a mouse model of breast cancer. Determination of TGIF1, E-cadherin and Twist1 protein levels in breast tumor tissue of mice in each group. Results: The tumor volume of mice in the TGIF1-overexpression group was significantly larger than that in the TGIF1-normal group and the TGIF1-silencing group, and the differences between the groups were statistically significant (P <0.05).The expression levels of TGIF1 and Twist1 protein in TGIF1-normal group, TGIF1-silencing group, and TGIF1-overexpression group were significantly higher than those in control group, and E-cadherin was significantly lower than that in control group. The differences between groups were statistically significant (P <0.05).The expression level of TGIF1 and Twist1 protein in TGIF1-silencing group was significantly lower than that in TGIF1-normal group, and E-cadherin was significantly higher than that in TGIF1-normal group (P < 0.05).The expression levels of TGIF1 and Twist1 proteins in the TGIF1-overexpression group were significantly higher than those in the TGIF1-normal group, and E-cadherin was significantly lower than that in the TGIF1-normal group. The differences between the groups were statistically significant (P <0.05). Pearson correlation analysis showed that the expression level of TGIF1 in breast cancer tissue was significantly negatively correlated with the expression level of E-cadherin protein, and was significantly positively correlated with the level of Twist1 protein (P <0.05). Conclusion: TGIF1 can affect the metastasis and invasion of breast cancer by regulating E-cadherin and Twist1 to interfere with the EMT pathway, which deserves further study.展开更多
TGF-β-induced factor homeobox 2(TGIF2)is a transcription regulator that plays essential roles in the regulation of development and cell fate decisions.Aberrant expression of TGIF family proteins has been observed in ...TGF-β-induced factor homeobox 2(TGIF2)is a transcription regulator that plays essential roles in the regulation of development and cell fate decisions.Aberrant expression of TGIF family proteins has been observed in several cancers,including ovarian,esophageal,and colorectal cancers.Here,we report that TGIF2 mediates the EGFR–RAS–ERK signaling pathway to enhance the stemness of lung adenocarcinoma(LUAD)cells and,therefore,promote the progression and metastasis of LUAD.We found that high TGIF2 expression was closely correlated with tumor growth,lymph node metastasis,and survival of patients with LUAD.Mice bearing TGIF2-silenced H1299 xenografts developed smaller tumors and fewer lung metastases.Importantly,silencing TGIF2 decreased the cancer stem cell(CSC)-like properties in A549 and H1299 cells.Furthermore,we identified that TGIF2 binding to the OCT4 promoter promotes its expression.In both LUAD cells and in vivo LUAD mouse models,we revealed that EGFR–RAS–ERK signaling phosphorylated TGIF2 and increased its stability,which was important for TGIF2-promoted LUAD stemness since phosphorylation-deficient TGIF2 mutants lost these functions.Thus,our study revealed that an important factor,TGIF2,bridges EGFR signaling to the CSC characteristics of LUAD cells,which can be utilized as an effective target for LUAD therapy.展开更多
SOX3 is a member of the Sox gene family implicated in brain formation and cognitive function. It is considered to be one of the earliest neural markers in vertebrates, playing a role in specifying neuronal fate. Recen...SOX3 is a member of the Sox gene family implicated in brain formation and cognitive function. It is considered to be one of the earliest neural markers in vertebrates, playing a role in specifying neuronal fate. Recently, we have established the first link between TALE （three- amino-acid loop extension） proteins, PBX1 （pre-B-cell leukemia homeobox 1） and MEIS1 （myeloid ecotropic viral integration site 1 homologue）, and the expression of the human SOX3 gene. Here we present the evidence that TGIF （TG-interacting factor） is an additional TALE superfamily member involved in the regulation of human SOX3 gene expression in NT2/D1 cells by direct interaction with the consensus binding site that is conserved in primate orthologue promoters. Functional analysis demonstrated that mutation of the TGIF binding site resulted in the activation of SOX3 promoter. TGIF overexpression downregulates SOX3 promoter activity and decreases endogenous SOX3 protein expression in both uninduced and retinoic acid （RA）-induced NT2/D1 ceils. Up to now, this is the first transcription factor identified as a negative regulator of SOX3 gene expression. The obtained results further underscore the significance of TALE proteins as important transcriptional regulators of SOX3 gene expression.展开更多
胆管癌是一种起病隐匿、侵袭性强、致死率高的原发性恶性肿瘤。多聚嘧啶区结合蛋白1(polypyrimidine tract-binding protein 1,PTBP1)已被报道,在多种类型肿瘤组织中异常高表达并参与癌症进展,但其在胆管癌中的作用仍未见报道。该研究...胆管癌是一种起病隐匿、侵袭性强、致死率高的原发性恶性肿瘤。多聚嘧啶区结合蛋白1(polypyrimidine tract-binding protein 1,PTBP1)已被报道,在多种类型肿瘤组织中异常高表达并参与癌症进展,但其在胆管癌中的作用仍未见报道。该研究旨在探讨PTBP1在胆管癌中的生物学功能,并初步解析其分子机制。本文利用公开的癌症基因组图谱(the cancer genome atlas,TCGA)数据,分析了胆管癌及癌旁组织中的PTBP1 mRNA表达水平。结果显示,PTBP1在胆管癌组织中的表达水平显著高于癌旁组织(P<0.05)。随后,在胆管癌细胞系RBE和HuH28中,通过CCK-8和细胞平板克隆实验,评价了PTBP1对胆管癌细胞生长能力的影响。结果显示,过表达PTBP1可显著促进胆管癌细胞的生长(P<0.01),而敲低PTBP1显著抑制胆管癌细胞的生长(P<0.001)。Transwell和Invasion实验结果显示,过表达PTBP1可显著促进胆管癌细胞的迁移和侵袭(P<0.001),而敲低PTBP1显著抑制胆管癌细胞的迁移和侵袭(P<0.001)。转录物组测序和通路富集分析结果显示,在胆管癌细胞中,敲低PTBP1后上调表达的基因显著富集于p53信号通路;而下调表达的基因显著富集于胆固醇代谢、Rho GTPase和TGF-β等信号通路。基于上述转录物组测序数据,本文还分析发现,敲低PTBP1可导致一系列基因发生异常的mRNA可变剪接事件,例如参与TGF-β调控的TGIF1及与p53活性相关的GNAS基因等。综上所述,PTBP1可能通过调控一系列基因的可变剪接而影响多个癌症相关的信号通路,从而促进胆管癌的进展。展开更多
基金the National Science Foundation of China (No. 305709)the National Basic Research Program of China (No. 2007CB947503).
文摘TG-interacting factors （TGIFs） belong to a family of TALE-homeodomain proteins including TGIF, TGIF2, and TGIF2LX/Y （TGIF2 like on X or Y chromosome） in human. They potentially play important functions in various tissues during development. Mutations in TGIF are frequently associated with malformation of forebrain and facial structures; TGIF2 proteins are over-expressed in many ovarian cancer cell lines; and TGIF2LX/Y are specifically expressed in adult testis. The molecular functions of these proteins have been investigated mostly in cultured cells. TGIF and TGIF2 have been found as transcriptional repressors that modulate TGF-beta signaling. However these findings are far from sufficient to explain their mutant phenotypes or expression patterns, and the functions of TGIF2LX/Y have never been reported. Here we use Drosophila as a model system to explore the functions of TGIF family proteins in vivo. We observed in fly tissues such as fat body, epithelia, and neuronal cells, that expressing human TGIF2 or human TGIF2LX generally inhibited cell growth in size and number. Co-expressing Drosophila Myc, Cyclin E, or human c-MycS partially rescued the growth inhibition induced by human TGIFs, whereas activated insulin pathway signaling did not. Taken together, we provide in vivo evidence for the potential functions of human TGIF2 and TGIF2LX in growth control. Additionally, we confirmed that Drosophila TGIFs are transcriptional activators by assaying their activities in spermatogenesis.
基金supported by the Independent Innovation in Jiangsu Province of China(CX(21)3119)the National Natural Science Foundation of China(31802035,31702095 and 31872324)+1 种基金the Jiangsu Natural Science Fund(BK20181221)Yangzhou Liangde Antibody Bio Tech.,China。
文摘Excess ammonia(NH_(3))in the circulation of dairy animals can reduce animal health and the quality of products for human consumption.To develop effective prevention and treatment methods,it is essential to examine the molecular mechanisms through which excess NH_(3) may affect the mammary gland.The present study used bovine mammary epithelial cells(BMECs)to evaluate the effects of exogenous NH_(4)Cl on the abundance of circular RNAs(circRNAs)using high-throughput sequencing.Among the identified circRNAs,circ02771 was the most significantly upregulated by exogenous NH_(4)Cl(P<0.05),with a fold change of 4.12.The results of the apoptosis and proliferation assays,transmission electron microscopy,H&E staining,and immunohistochemistry revealed that circ02771 increased apoptosis and inflammation.A double luciferase reporter assay revealed that circ02771 targeted miR-194b,and the overexpression of circ02771(pcDNA-circ02771)reduced(P<0.05)the expression of miR-194b and led to apoptosis and inflammation.Circ02771 also enhanced the expression of transforming growth factor beta-induced factor homeobox 1(TGIF1),which is a target gene of miR-194b.Overall,this study suggests that the circ02771/miR-194b/TGIF1 axis plays a role in mediating the effects of NH_(4)Cl on BMECs.Therefore,this axis provides a novel target to help control hazards within the mammary gland from high circulating NH_(4)Cl levels.
基金This research was supported by the National Natural Science Foundation of China(No.31802263)Outstanding Young Talent Project of the Beijing Municipal Party Committee Organization Department(No.2018000020124G081).
文摘Endometritis affects the reproductive capacity of dairy cows and leads to serious economic losses in dairy farming.Clarification of the pathogenesis of endometritis is necessary to improve the reproductive efficiency of dairy cows.Exosomes and their miRNAs have been proven to play an important role in inflammatory regulation.Exosomal miR-218 is a differentially expressed miRNA found in endometrial epithelial cells(EECs)under endometrial inflammation.Therefore,we investigated the expression of miR-218 in the uterine tissue of dairy cows,lipopolysaccharide(LPS)treated EECs,exosomal vesicles,and regulation of exosomal miR-218 by targeting TGIF-2 inducible factor homology frame 2(TGIF2)/transforming growth factor-beta(TGF-β).The expression of miR-218 was suppressed in inflammatory uterine tissues and LPS treated EECs.The expression of TGIF2 and TGF-βin inflammatory uterine tissues and LPS treated EECs was significantly higher than those in healthy uterine tissues and EECs(p<0.01).Interestingly,miR-218 derived from donor cells was found to regulate the expression of the target gene TGIF2 in recipient cells through the fusion of exosomes.Concurrently,the expression of its target gene TGIF2 was also suppressed by miR-218 in donor cells resulting in fewer TGIF2 being transported into recipient cells with exosomal fusion.This may be a novel mechanism of miRNAs-mediated regulation and provides a new reference for analyzing the pathogenesis of endometritis in dairy cows.
基金National natural science foundation of Gansu province(160RJZA170)
文摘Objective: To observe the effect of the expression of transforming growth factor β-homologous inducible factor 1 (TGIF1) on the expression of epithelial cadherin (E-cadherin) and human Twist-related protein 1 (Twist1) in breast cancer cells. Methods: Total of twenty-four 6-week-old female SPF Balb/c mice were randomly divided into a control group, a TGIF1-silencing group, a TGIF1-normal group, and a TGIF1-overexpression group. In the TGIF1-silencing group, 4T1 breast cancer cells were interfered by lentivirus shRNA (H) lentiviral particles (sc-36659-v) to construct a breast cancer model. TGIF1-normal group used breast cancer cells (4T1) to construct a mouse model of breast cancer. And the TGIF1-overexpression group used 4T1 breast cancer cells with TGIF1 overexpression to establish a mouse model of breast cancer. Determination of TGIF1, E-cadherin and Twist1 protein levels in breast tumor tissue of mice in each group. Results: The tumor volume of mice in the TGIF1-overexpression group was significantly larger than that in the TGIF1-normal group and the TGIF1-silencing group, and the differences between the groups were statistically significant (P <0.05).The expression levels of TGIF1 and Twist1 protein in TGIF1-normal group, TGIF1-silencing group, and TGIF1-overexpression group were significantly higher than those in control group, and E-cadherin was significantly lower than that in control group. The differences between groups were statistically significant (P <0.05).The expression level of TGIF1 and Twist1 protein in TGIF1-silencing group was significantly lower than that in TGIF1-normal group, and E-cadherin was significantly higher than that in TGIF1-normal group (P < 0.05).The expression levels of TGIF1 and Twist1 proteins in the TGIF1-overexpression group were significantly higher than those in the TGIF1-normal group, and E-cadherin was significantly lower than that in the TGIF1-normal group. The differences between the groups were statistically significant (P <0.05). Pearson correlation analysis showed that the expression level of TGIF1 in breast cancer tissue was significantly negatively correlated with the expression level of E-cadherin protein, and was significantly positively correlated with the level of Twist1 protein (P <0.05). Conclusion: TGIF1 can affect the metastasis and invasion of breast cancer by regulating E-cadherin and Twist1 to interfere with the EMT pathway, which deserves further study.
基金This work was supported by the National Natural Science Foundation of China(No.81772974 to Y.Shi,No.1672914 to Y.Luo,and No.81972795 to C.Chen).
文摘TGF-β-induced factor homeobox 2(TGIF2)is a transcription regulator that plays essential roles in the regulation of development and cell fate decisions.Aberrant expression of TGIF family proteins has been observed in several cancers,including ovarian,esophageal,and colorectal cancers.Here,we report that TGIF2 mediates the EGFR–RAS–ERK signaling pathway to enhance the stemness of lung adenocarcinoma(LUAD)cells and,therefore,promote the progression and metastasis of LUAD.We found that high TGIF2 expression was closely correlated with tumor growth,lymph node metastasis,and survival of patients with LUAD.Mice bearing TGIF2-silenced H1299 xenografts developed smaller tumors and fewer lung metastases.Importantly,silencing TGIF2 decreased the cancer stem cell(CSC)-like properties in A549 and H1299 cells.Furthermore,we identified that TGIF2 binding to the OCT4 promoter promotes its expression.In both LUAD cells and in vivo LUAD mouse models,we revealed that EGFR–RAS–ERK signaling phosphorylated TGIF2 and increased its stability,which was important for TGIF2-promoted LUAD stemness since phosphorylation-deficient TGIF2 mutants lost these functions.Thus,our study revealed that an important factor,TGIF2,bridges EGFR signaling to the CSC characteristics of LUAD cells,which can be utilized as an effective target for LUAD therapy.
基金supported by the Ministry of Education and Science,Republic of Serbia(Nos.143028 and 173051)
文摘SOX3 is a member of the Sox gene family implicated in brain formation and cognitive function. It is considered to be one of the earliest neural markers in vertebrates, playing a role in specifying neuronal fate. Recently, we have established the first link between TALE （three- amino-acid loop extension） proteins, PBX1 （pre-B-cell leukemia homeobox 1） and MEIS1 （myeloid ecotropic viral integration site 1 homologue）, and the expression of the human SOX3 gene. Here we present the evidence that TGIF （TG-interacting factor） is an additional TALE superfamily member involved in the regulation of human SOX3 gene expression in NT2/D1 cells by direct interaction with the consensus binding site that is conserved in primate orthologue promoters. Functional analysis demonstrated that mutation of the TGIF binding site resulted in the activation of SOX3 promoter. TGIF overexpression downregulates SOX3 promoter activity and decreases endogenous SOX3 protein expression in both uninduced and retinoic acid （RA）-induced NT2/D1 ceils. Up to now, this is the first transcription factor identified as a negative regulator of SOX3 gene expression. The obtained results further underscore the significance of TALE proteins as important transcriptional regulators of SOX3 gene expression.
文摘胆管癌是一种起病隐匿、侵袭性强、致死率高的原发性恶性肿瘤。多聚嘧啶区结合蛋白1(polypyrimidine tract-binding protein 1,PTBP1)已被报道,在多种类型肿瘤组织中异常高表达并参与癌症进展,但其在胆管癌中的作用仍未见报道。该研究旨在探讨PTBP1在胆管癌中的生物学功能,并初步解析其分子机制。本文利用公开的癌症基因组图谱(the cancer genome atlas,TCGA)数据,分析了胆管癌及癌旁组织中的PTBP1 mRNA表达水平。结果显示,PTBP1在胆管癌组织中的表达水平显著高于癌旁组织(P<0.05)。随后,在胆管癌细胞系RBE和HuH28中,通过CCK-8和细胞平板克隆实验,评价了PTBP1对胆管癌细胞生长能力的影响。结果显示,过表达PTBP1可显著促进胆管癌细胞的生长(P<0.01),而敲低PTBP1显著抑制胆管癌细胞的生长(P<0.001)。Transwell和Invasion实验结果显示,过表达PTBP1可显著促进胆管癌细胞的迁移和侵袭(P<0.001),而敲低PTBP1显著抑制胆管癌细胞的迁移和侵袭(P<0.001)。转录物组测序和通路富集分析结果显示,在胆管癌细胞中,敲低PTBP1后上调表达的基因显著富集于p53信号通路;而下调表达的基因显著富集于胆固醇代谢、Rho GTPase和TGF-β等信号通路。基于上述转录物组测序数据,本文还分析发现,敲低PTBP1可导致一系列基因发生异常的mRNA可变剪接事件,例如参与TGF-β调控的TGIF1及与p53活性相关的GNAS基因等。综上所述,PTBP1可能通过调控一系列基因的可变剪接而影响多个癌症相关的信号通路,从而促进胆管癌的进展。
