Objective:To investigate the effects and possible mechanisms of action of Curcuma wenyujin Y.H.Chen et C.Ling n-Butyl alcohol extract(CWNAE)on repression of human gastric cancer(GC)AGS cell invasion induced by co-cult...Objective:To investigate the effects and possible mechanisms of action of Curcuma wenyujin Y.H.Chen et C.Ling n-Butyl alcohol extract(CWNAE)on repression of human gastric cancer(GC)AGS cell invasion induced by co-culturing with Helicobacter pylori(HP).Methods:AGS cells were cultured with HP of positive or negative cytotoxin-associated gene A(Cag A)and vacuolating cytotoxin gene A(Vac A)expression(Cag A+/-or Vac A+/-)and divided into 5 group.Group A was cultured without HP as a control,Group B with HPCagA+VacA+,Group C with HPCagA-VacA-,Group D with HPCagA+VacA+and CWNAE,and Group E with HPCag A-Vac Aand CWNAE.Methylthiazolyldiphenyl-tetrazolium bromide(MTT)and tumor invasion assays,examinations of morphology and ultramicroscopic structures,quantitative real-time polymerase chain reaction and Western blots were performed to measure the effects and uncover the mechanisms behind these effects of HPCagA+VacA+and CWNAE on the epithelial-mesenchymal transition(EMT)of AGS cells.Results:The 10%inhibitory concentration of CWNAE against AGS cells after a 48 h incubation was 19.73±1.30μg/m L.More AGS cells were elongated after co-culturing with HPCagA+VacA+than after culturing with HPCagA-VacA-.In tumor invasion assays,HPCagA+VacA+significantly enhanced the invasiveness of AGS cells compared to the other experimental groups(all P value<0.05),and this effect was inhibited by CWNAE.Treatment with CWNAE normalized tight junctions and reduced the number of pseudopodia of AGS cells co-cultured with HPCagA+VacA+.HPCagA+VacA+up-regulated zincfinger ebox binding homeobox 1(ZEB1)in AGS cells after co-culturing for 24 h.Expression of caudal type homeobox transcription factor(CDX-2)and claudin-2 was significantly increased by HPCagA+VacA+(P<0.05),but not by HPCagA-VacA-.Conclusions:HPCagA+VacA+promoted the invasiveness of AGS cells through up-regulation of ZEB1 transcription and claudin-2 and CDX-2 expression.CWNAE inhibited these effects of HPCagA+VacA+on AGS cells by down-regulating ZEB1 transcription,and CDX-2 and claudin-2 expression.展开更多
目的对孤立性心房颤动(AF)患者进行垂体同型框2(PITX2)基因的突变扫描,并分析突变患者的相应临床特点。方法对160例孤立性AF患者和200名健康者的PITX2基因进行突变扫描。采用聚合酶链反应(PCR)扩增PITX2基因,通过单链构象多态性技术筛...目的对孤立性心房颤动(AF)患者进行垂体同型框2(PITX2)基因的突变扫描,并分析突变患者的相应临床特点。方法对160例孤立性AF患者和200名健康者的PITX2基因进行突变扫描。采用聚合酶链反应(PCR)扩增PITX2基因,通过单链构象多态性技术筛查突变,对发现异常的个体采用双脱氧核苷链末端合成终止法进行直接测序。采用Sequence scanner v 1.0软件分析测序图。结果在2例永久性AF患者中发现位于内含子区域的新改变g.766C>A、位于5’UTR区域的新改变g.8310C>T(c.-150C>T)。这两例携带者对药物及电复律治疗的反应较差,接受环肺静脉消融手术后,随访1年未再复发。200名健康者中未发现此改变。结论首次在中国孤立性AF患者中发现影响肺静脉肌袖发育基因PITX2的非编码区改变,其可能为我们进一步理解肺静脉肌袖在AF的发病过程中的作用提供新的启示。展开更多
基金Supported by the National Natural Science Foundation of China(No.81202822)Fund of Zhejiang Province Administration of Traditional Chinese Medicine,China(No.2016ZA092/2017ZKL008)Key Laboratory of Pathology and Physiology of Digestive Tract Diseases in Zhejiang Province,China
文摘Objective:To investigate the effects and possible mechanisms of action of Curcuma wenyujin Y.H.Chen et C.Ling n-Butyl alcohol extract(CWNAE)on repression of human gastric cancer(GC)AGS cell invasion induced by co-culturing with Helicobacter pylori(HP).Methods:AGS cells were cultured with HP of positive or negative cytotoxin-associated gene A(Cag A)and vacuolating cytotoxin gene A(Vac A)expression(Cag A+/-or Vac A+/-)and divided into 5 group.Group A was cultured without HP as a control,Group B with HPCagA+VacA+,Group C with HPCagA-VacA-,Group D with HPCagA+VacA+and CWNAE,and Group E with HPCag A-Vac Aand CWNAE.Methylthiazolyldiphenyl-tetrazolium bromide(MTT)and tumor invasion assays,examinations of morphology and ultramicroscopic structures,quantitative real-time polymerase chain reaction and Western blots were performed to measure the effects and uncover the mechanisms behind these effects of HPCagA+VacA+and CWNAE on the epithelial-mesenchymal transition(EMT)of AGS cells.Results:The 10%inhibitory concentration of CWNAE against AGS cells after a 48 h incubation was 19.73±1.30μg/m L.More AGS cells were elongated after co-culturing with HPCagA+VacA+than after culturing with HPCagA-VacA-.In tumor invasion assays,HPCagA+VacA+significantly enhanced the invasiveness of AGS cells compared to the other experimental groups(all P value<0.05),and this effect was inhibited by CWNAE.Treatment with CWNAE normalized tight junctions and reduced the number of pseudopodia of AGS cells co-cultured with HPCagA+VacA+.HPCagA+VacA+up-regulated zincfinger ebox binding homeobox 1(ZEB1)in AGS cells after co-culturing for 24 h.Expression of caudal type homeobox transcription factor(CDX-2)and claudin-2 was significantly increased by HPCagA+VacA+(P<0.05),but not by HPCagA-VacA-.Conclusions:HPCagA+VacA+promoted the invasiveness of AGS cells through up-regulation of ZEB1 transcription and claudin-2 and CDX-2 expression.CWNAE inhibited these effects of HPCagA+VacA+on AGS cells by down-regulating ZEB1 transcription,and CDX-2 and claudin-2 expression.
文摘目的对孤立性心房颤动(AF)患者进行垂体同型框2(PITX2)基因的突变扫描,并分析突变患者的相应临床特点。方法对160例孤立性AF患者和200名健康者的PITX2基因进行突变扫描。采用聚合酶链反应(PCR)扩增PITX2基因,通过单链构象多态性技术筛查突变,对发现异常的个体采用双脱氧核苷链末端合成终止法进行直接测序。采用Sequence scanner v 1.0软件分析测序图。结果在2例永久性AF患者中发现位于内含子区域的新改变g.766C>A、位于5’UTR区域的新改变g.8310C>T(c.-150C>T)。这两例携带者对药物及电复律治疗的反应较差,接受环肺静脉消融手术后,随访1年未再复发。200名健康者中未发现此改变。结论首次在中国孤立性AF患者中发现影响肺静脉肌袖发育基因PITX2的非编码区改变,其可能为我们进一步理解肺静脉肌袖在AF的发病过程中的作用提供新的启示。
