AIM: To explore the expression of transient receptor potential vanilloid 4(TRPV4) and its physiological meaning in mouse and rat gastric epithelia. METHODS: RT-PCR and immunochemistry were used to detect TRPV4 m RNA a...AIM: To explore the expression of transient receptor potential vanilloid 4(TRPV4) and its physiological meaning in mouse and rat gastric epithelia. METHODS: RT-PCR and immunochemistry were used to detect TRPV4 m RNA and protein expression in mouse stomach and a rat normal gastric epithelial cell line(RGE1-01), while Ca2+-imaging and electrophysiology were used to evaluate TRPV4 channel activity. ATP release was measured by a luciferin-luciferase assay. Gastric emptying was also compared between WT and TRPV4 knockout mice. RESULTS: TRPV4 m RNA and protein were detected in mouse tissues and RGE1-01 cells. A TRPV4-specific agonist(GSK1016790A) increased intracellular Ca2+ concentrations and/or evoked TRPV4-like current activities in WT mouse gastric epithelial cells andRGE1-01 cells, but not TRPV4 KO cells. GSK1016790 A or mechanical stimuli induced ATP release from RGE1-01 cells while TRPV4 knockout mice displayed delayed gastric emptying in vivo. CONCLUSION: TRPV4 is expressed in mouse and rat gastric epithelium and contributes to ATP release and gastric emptying.展开更多
多药耐药相关蛋白4(multidrug resistance-associated protein 4,MRP4)是一种能够利用ATP水解产生的能量转运多种物质的六重跨膜蛋白。本研究应用Expasy、Jpred4、UniProt-KB、NCBI等公共数据库和在线软件分析其理化性质、蛋白结构、结...多药耐药相关蛋白4(multidrug resistance-associated protein 4,MRP4)是一种能够利用ATP水解产生的能量转运多种物质的六重跨膜蛋白。本研究应用Expasy、Jpred4、UniProt-KB、NCBI等公共数据库和在线软件分析其理化性质、蛋白结构、结构域、亚细胞定位、互作蛋白以及功能等信息,为其在多种生理病理过程中的作用与机制研究奠定基础。MRP4蛋白含有1325个氨基酸,相对分子质量约为149526.77,理论等电点为8.41。以α-螺旋为主,占比58.42%。含有2个跨膜结构域和2个ATP结合盒以及8个保守结构域。表观遗传学修饰位点56个,其中磷酸化位点27个,泛素化位点27个,乙酰化位点2个。MRP4蛋白分布于质膜上,具有ATP酶和跨膜转运蛋白活性,参与内外源性物质运输、前列腺素分泌、纤毛组装、信号转导运输等生命过程。本研究系统性梳理并整合了MRP4蛋白结构及功能等信息,为深入研究其在机体生命活动及疾病发生中的作用与机制奠定理论依据。展开更多
目的探索在人胆囊癌细胞系中是否存在具有干细胞特性的SP细胞（side population cells）以及SP细胞、非SP细胞、GBCSD细胞系之间在耐药基因ABCG2和胚胎干细胞表面标志Oct-4表达方面的差异性。方法采用FACS（流式激活细胞分选）技术分选...目的探索在人胆囊癌细胞系中是否存在具有干细胞特性的SP细胞（side population cells）以及SP细胞、非SP细胞、GBCSD细胞系之间在耐药基因ABCG2和胚胎干细胞表面标志Oct-4表达方面的差异性。方法采用FACS（流式激活细胞分选）技术分选出人胆囊癌SP细胞和非SP细胞，通过RTPCR、Western blot以及流式技术来检测SP细胞、非SP细胞以及GBC—SD细胞系在ABCG2和Oct-4表达方面的差异情况。结果人胆囊癌GBC—SD细胞系中存在着具有干细胞潜能的SP细胞，其所占的比例为（0．64±0．08）％。并且ABCG2在胆囊癌SP细胞中呈现出高表达的状态[（89．56±3．86）%]．在非SP细胞中几乎不表达[（1．32±0．49）％]，两者之间的差异具有统计学意义（P〈0．05）．在GBCSD细胞系中呈弱表达[（12．37±1．61）％]。Oct-4在三种细胞中的表达分别为：（94．87±1．40）％、（88．16±2．34）％、（90．17±1．61）％，三者之间差异没有统计学意义（P〉0．05）。结论人胆囊癌的起源与肿瘤干细胞密切相关并且在胆囊癌细胞系中存在着具有干细胞特性并且高表达耐药基因ABCG2的人胆囊癌SP细胞。展开更多
基金Supported by Grants from the University of Toyama and JSPS KAKENHI to Mihara H,No.26870214
文摘AIM: To explore the expression of transient receptor potential vanilloid 4(TRPV4) and its physiological meaning in mouse and rat gastric epithelia. METHODS: RT-PCR and immunochemistry were used to detect TRPV4 m RNA and protein expression in mouse stomach and a rat normal gastric epithelial cell line(RGE1-01), while Ca2+-imaging and electrophysiology were used to evaluate TRPV4 channel activity. ATP release was measured by a luciferin-luciferase assay. Gastric emptying was also compared between WT and TRPV4 knockout mice. RESULTS: TRPV4 m RNA and protein were detected in mouse tissues and RGE1-01 cells. A TRPV4-specific agonist(GSK1016790A) increased intracellular Ca2+ concentrations and/or evoked TRPV4-like current activities in WT mouse gastric epithelial cells andRGE1-01 cells, but not TRPV4 KO cells. GSK1016790 A or mechanical stimuli induced ATP release from RGE1-01 cells while TRPV4 knockout mice displayed delayed gastric emptying in vivo. CONCLUSION: TRPV4 is expressed in mouse and rat gastric epithelium and contributes to ATP release and gastric emptying.
文摘多药耐药相关蛋白4(multidrug resistance-associated protein 4,MRP4)是一种能够利用ATP水解产生的能量转运多种物质的六重跨膜蛋白。本研究应用Expasy、Jpred4、UniProt-KB、NCBI等公共数据库和在线软件分析其理化性质、蛋白结构、结构域、亚细胞定位、互作蛋白以及功能等信息,为其在多种生理病理过程中的作用与机制研究奠定基础。MRP4蛋白含有1325个氨基酸,相对分子质量约为149526.77,理论等电点为8.41。以α-螺旋为主,占比58.42%。含有2个跨膜结构域和2个ATP结合盒以及8个保守结构域。表观遗传学修饰位点56个,其中磷酸化位点27个,泛素化位点27个,乙酰化位点2个。MRP4蛋白分布于质膜上,具有ATP酶和跨膜转运蛋白活性,参与内外源性物质运输、前列腺素分泌、纤毛组装、信号转导运输等生命过程。本研究系统性梳理并整合了MRP4蛋白结构及功能等信息,为深入研究其在机体生命活动及疾病发生中的作用与机制奠定理论依据。
文摘目的探索在人胆囊癌细胞系中是否存在具有干细胞特性的SP细胞（side population cells）以及SP细胞、非SP细胞、GBCSD细胞系之间在耐药基因ABCG2和胚胎干细胞表面标志Oct-4表达方面的差异性。方法采用FACS（流式激活细胞分选）技术分选出人胆囊癌SP细胞和非SP细胞，通过RTPCR、Western blot以及流式技术来检测SP细胞、非SP细胞以及GBC—SD细胞系在ABCG2和Oct-4表达方面的差异情况。结果人胆囊癌GBC—SD细胞系中存在着具有干细胞潜能的SP细胞，其所占的比例为（0．64±0．08）％。并且ABCG2在胆囊癌SP细胞中呈现出高表达的状态[（89．56±3．86）%]．在非SP细胞中几乎不表达[（1．32±0．49）％]，两者之间的差异具有统计学意义（P〈0．05）．在GBCSD细胞系中呈弱表达[（12．37±1．61）％]。Oct-4在三种细胞中的表达分别为：（94．87±1．40）％、（88．16±2．34）％、（90．17±1．61）％，三者之间差异没有统计学意义（P〉0．05）。结论人胆囊癌的起源与肿瘤干细胞密切相关并且在胆囊癌细胞系中存在着具有干细胞特性并且高表达耐药基因ABCG2的人胆囊癌SP细胞。
