目的利用小分子干扰RNA(small interfering RNA,s i R N A)技术探究体外沉默K rüp p e l样因子17(Krüppel like factor 17,KLF17)的基因表达对结直肠癌细胞株SW480细胞增殖和迁移的影响,为抑制结直肠癌复发转移奠定理论基础....目的利用小分子干扰RNA(small interfering RNA,s i R N A)技术探究体外沉默K rüp p e l样因子17(Krüppel like factor 17,KLF17)的基因表达对结直肠癌细胞株SW480细胞增殖和迁移的影响,为抑制结直肠癌复发转移奠定理论基础.方法使用基因重组方法构建KLF17的si RNA真核质粒表达载体,用电转染法转染至人结直肠癌SW480细胞中.使用荧光定量PCR检测KLF17、E-钙黏素(E-cadherin)和波形蛋白(Vimentin)的基因表达水平;用Western blot检测KLF17的蛋白表达以及细胞转移相关蛋白E-cadherin和Vimentin的表达.用四甲基偶氮唑蓝法检测细胞增殖活性.结果与对照组相比,转染si RNA后的SW480细胞培养48 h后增殖能力受到显著抑制,且细胞形态由圆形或多边形变成梭形并向外伸出多个细胞突起;转染si RNA后KLF17和E-cadherin蛋白表达以及基因表达水平显著降低,Vimentin的蛋白表达水平和基因表达水平显著升高.结论抑制KLF17的表达可以通过诱导结直肠癌细胞发生上皮-间质转化而促进其细胞的增殖以及转移.展开更多
目的:探讨Kruppel样因子17(Kruppel-like factor 17,KLF17)、转录辅助因子退变样蛋白4(vestigial like family member 4,VGLL4)在子宫内膜样腺癌组织中的表达,了解二者在不同病理类型的表达差异及其与临床病理特征之间的联系。方法:选择...目的:探讨Kruppel样因子17(Kruppel-like factor 17,KLF17)、转录辅助因子退变样蛋白4(vestigial like family member 4,VGLL4)在子宫内膜样腺癌组织中的表达,了解二者在不同病理类型的表达差异及其与临床病理特征之间的联系。方法:选择2015年6月-2020年4月清远市妇幼保健院妇产科收集经病理确诊的标本共91例,子宫内膜样腺癌组织(41例),子宫内膜单纯性增生组织(28例),正常子宫内膜组织(22例)。运用免疫组化法检测KLF17、VGLL4的表达水平,分析KLF17、VGLL4与临床病理特征的关系。结果:KLF17在子宫内膜样腺癌及子宫内膜单纯性增生的阳性表达率分别为80.5%、25.0%,高于正常子宫内膜的18.2%,差异有统计学意义(P<0.05)。VGLL4在子宫内膜样腺癌的阳性表达率24.4%,低于子宫内膜单纯性增生、正常子宫内膜的71.4%、77.3%,差异有统计学意义(P<0.05)。KLF17与VGLL4在子宫内膜样腺癌中的表达,二者之间存在负相关(r=-0.437,P<0.05)。在人子宫内膜样腺癌组织中提示VGLL4表达与子宫内膜样腺癌分化程度存在正相关(r=0.346,P<0.05)。KLF17在子宫内膜样腺癌中的表达与患者年龄、FIGO分期、肌层浸润深度、分化程度、淋巴结转移无关(P>0.05)。VGLL4在子宫内膜样腺癌中的表达与患者年龄、FIGO分期、肌层浸润深度及淋巴结转移等无关(P>0.05),而与分化程度有相关性,分化程度越高,表达越强(P<0.05)。结论:在子宫内膜样腺癌组织中,KLF17表达上调和VGLL4表达下调,二者可能与内膜病变的发生发展有关。展开更多
The pluripotent state of embryonic stem cells(ESCs)is regulated by a sophisticated network of transcription factors.High expression of KLF17 has recently been identified as a hallmark of naive state of human ESCs(h ES...The pluripotent state of embryonic stem cells(ESCs)is regulated by a sophisticated network of transcription factors.High expression of KLF17 has recently been identified as a hallmark of naive state of human ESCs(h ESCs).However,the functional role of KLF17 in naive state is not clear.Here,by employing various gain and loss-of-function approaches,we demonstrate that KLF17 is essential for the maintenance of naive state and promotes the primed to naive state transition in h ESCs.Mechanistically,we identify MAPK3 and ZIC2 as two direct targets repressed by KLF17.Overexpression of MAPK3 or ZIC2 partially blocks KLF17 from promoting the naive pluripotency.Furthermore,we find that human and mouse homologs of KLF17 retain conserved functions in promoting naive pluripotency of both species.Finally,we show that Klf17 may be essential for early embryo development in mouse.These findings demonstrate the important and conserved function of KLF17 in promoting naive pluripotency and reveal two essential transcriptional targets of KLF17 that underlie its function.展开更多
文摘目的利用小分子干扰RNA(small interfering RNA,s i R N A)技术探究体外沉默K rüp p e l样因子17(Krüppel like factor 17,KLF17)的基因表达对结直肠癌细胞株SW480细胞增殖和迁移的影响,为抑制结直肠癌复发转移奠定理论基础.方法使用基因重组方法构建KLF17的si RNA真核质粒表达载体,用电转染法转染至人结直肠癌SW480细胞中.使用荧光定量PCR检测KLF17、E-钙黏素(E-cadherin)和波形蛋白(Vimentin)的基因表达水平;用Western blot检测KLF17的蛋白表达以及细胞转移相关蛋白E-cadherin和Vimentin的表达.用四甲基偶氮唑蓝法检测细胞增殖活性.结果与对照组相比,转染si RNA后的SW480细胞培养48 h后增殖能力受到显著抑制,且细胞形态由圆形或多边形变成梭形并向外伸出多个细胞突起;转染si RNA后KLF17和E-cadherin蛋白表达以及基因表达水平显著降低,Vimentin的蛋白表达水平和基因表达水平显著升高.结论抑制KLF17的表达可以通过诱导结直肠癌细胞发生上皮-间质转化而促进其细胞的增殖以及转移.
文摘目的:探讨Kruppel样因子17(Kruppel-like factor 17,KLF17)、转录辅助因子退变样蛋白4(vestigial like family member 4,VGLL4)在子宫内膜样腺癌组织中的表达,了解二者在不同病理类型的表达差异及其与临床病理特征之间的联系。方法:选择2015年6月-2020年4月清远市妇幼保健院妇产科收集经病理确诊的标本共91例,子宫内膜样腺癌组织(41例),子宫内膜单纯性增生组织(28例),正常子宫内膜组织(22例)。运用免疫组化法检测KLF17、VGLL4的表达水平,分析KLF17、VGLL4与临床病理特征的关系。结果:KLF17在子宫内膜样腺癌及子宫内膜单纯性增生的阳性表达率分别为80.5%、25.0%,高于正常子宫内膜的18.2%,差异有统计学意义(P<0.05)。VGLL4在子宫内膜样腺癌的阳性表达率24.4%,低于子宫内膜单纯性增生、正常子宫内膜的71.4%、77.3%,差异有统计学意义(P<0.05)。KLF17与VGLL4在子宫内膜样腺癌中的表达,二者之间存在负相关(r=-0.437,P<0.05)。在人子宫内膜样腺癌组织中提示VGLL4表达与子宫内膜样腺癌分化程度存在正相关(r=0.346,P<0.05)。KLF17在子宫内膜样腺癌中的表达与患者年龄、FIGO分期、肌层浸润深度、分化程度、淋巴结转移无关(P>0.05)。VGLL4在子宫内膜样腺癌中的表达与患者年龄、FIGO分期、肌层浸润深度及淋巴结转移等无关(P>0.05),而与分化程度有相关性,分化程度越高,表达越强(P<0.05)。结论:在子宫内膜样腺癌组织中,KLF17表达上调和VGLL4表达下调,二者可能与内膜病变的发生发展有关。
基金supported by the National Key Research and Development Program of China(2018YFA0107601 and2021YFA0100200)the National Natural Science Foundation of China(91940302,32130017,82070294 and 32025007)。
文摘The pluripotent state of embryonic stem cells(ESCs)is regulated by a sophisticated network of transcription factors.High expression of KLF17 has recently been identified as a hallmark of naive state of human ESCs(h ESCs).However,the functional role of KLF17 in naive state is not clear.Here,by employing various gain and loss-of-function approaches,we demonstrate that KLF17 is essential for the maintenance of naive state and promotes the primed to naive state transition in h ESCs.Mechanistically,we identify MAPK3 and ZIC2 as two direct targets repressed by KLF17.Overexpression of MAPK3 or ZIC2 partially blocks KLF17 from promoting the naive pluripotency.Furthermore,we find that human and mouse homologs of KLF17 retain conserved functions in promoting naive pluripotency of both species.Finally,we show that Klf17 may be essential for early embryo development in mouse.These findings demonstrate the important and conserved function of KLF17 in promoting naive pluripotency and reveal two essential transcriptional targets of KLF17 that underlie its function.
