目的:探讨体外沉默Kruppel样因子4(Kruppel like factor 4,KLF4)基因的表达对食管癌KYSE140细胞增殖及迁移的影响。方法:Western blotting法检测人食管癌细胞株KYSE140、KYSE150、EC109及EC9706及食管永生化细胞NE3中KLF4蛋白的表达,化...目的:探讨体外沉默Kruppel样因子4(Kruppel like factor 4,KLF4)基因的表达对食管癌KYSE140细胞增殖及迁移的影响。方法:Western blotting法检测人食管癌细胞株KYSE140、KYSE150、EC109及EC9706及食管永生化细胞NE3中KLF4蛋白的表达,化学合成2对靶向KLF4的siRNA(KLF4-siRNA1,KLF4-siRNA2),并设对照siRNA(Ctrl-siRNA),分别体外转染至高表达KLF4的食管癌KYSE140细胞中,形成KLF4-siRNA1-KYSE140、KLF4-siRNA2-KYSE140及Ctrl-siRNA-KYSE140细胞,通过MTT实验、Transwell实验分别检测转染后食管癌KYSE140细胞的增殖及迁移。结果:食管癌细胞株KYSE140中KLF4蛋白的表达明显高于KYSE150、EC109及EC9706细胞株[(5.62±0.02)vs(1.71±0.23)、(3.24±0.35)、(3.16±0.41),均P<0.05]。KLF4-siRNA1-KYSE140、KLF4-siRNA2-KYSE140与Ctrl-siRNA-KYSE140细胞相比,KLF4蛋白表达明显降低[(0.49±0.18)、(0.32±0.09)vs(0.98±0.19),均P<0.05],细胞增殖能力明显增高[(1.2±0.8)、(1.4±0.1)vs(0.6±0.1),均P<0.05],迁移细胞数量也明显增加[(780±22)、(475±25)vs(83±17)个,P<0.05]。结论:KLF4在人食管癌细胞的增殖和迁移过程中起着负调控作用。展开更多
Salidroside is extensively used as a herbal medicine worldwide, and it has been shown to protect against disruption of endothelial homeostasis and act as an anti-aging agent. The present study aimed to investigate the...Salidroside is extensively used as a herbal medicine worldwide, and it has been shown to protect against disruption of endothelial homeostasis and act as an anti-aging agent. The present study aimed to investigate the ameliorative effects of salidroside on homocysteine (Hcy)-induced cell senescence in human umbilical vein endothelial cells (HUVECs) that were mediated via inhibition of Krüppel-like factor 4 (KLF4). An endothelial cell senescence model was induced by Hcy. The cell viability, activities of telomerase and lactate dehydrogenase (LDH), and the level of reactive oxygen species were determined using commercial kits. The expression levels of KLF4, p53 and p21 were determined via western blot analysis, whereas the mRNA expression levels of KLF4 were detected by reverse transcription-quantitative PCR. Small interfering RNA-mediated knockdown of KLF4 was found to reverse Hcy-induced cell senescence. Hcy treatment led to an accelerated cell senescence, as evidenced by decreases in both cell viability and telomerase activity, whereas increases were noted in the leakage of LDH and the level of reactive oxygen species, in addition to an up-regulation of the protein levels of p53 and p21, and up-regulation of KLF4 at both the mRNA and protein level. Treatment with salidroside ameliorated Hcy-induced cell senescence in a dose-dependent manner. Taken together, these results suggested that Hcy may induce cell senescence through upregulation of KLF4, and this may be reversed by treatment with salidroside. Therefore, salidroside was shown to inhibit Hcy-induced cell senescence through KLF4 inhibition.展开更多
Kruppel-like factor 4（Klf4） is a zinc finger transcription factor and plays crucial roles in Xenopus embryogenesis.However, its regulation during embryogenesis is still unclear. Here, we report that Tcf711, a key do...Kruppel-like factor 4（Klf4） is a zinc finger transcription factor and plays crucial roles in Xenopus embryogenesis.However, its regulation during embryogenesis is still unclear. Here, we report that Tcf711, a key downstream transducer of the Wnt signaling pathway, could promote Klf4 transcription and stimulate Klf4 promoter activity in early Xenopus embryos. Furthermore, cycloheximide treatment showed a direct effect on Klf4 transcription facilitated by Tcf711. Moreover, the dominant negative form of Tcf711（dnTcf711）, which lacks N-terminus of the β-catenin binding motif, could still activate Klf4 transcription, suggesting that this regulation is Wnt/β-catenin independent.Taken together, our results demonstrate that Tcf711 lies upstream of Klf4 to maintain its expression level during Xenopus embryogenesis.展开更多
文摘目的:探讨体外沉默Kruppel样因子4(Kruppel like factor 4,KLF4)基因的表达对食管癌KYSE140细胞增殖及迁移的影响。方法:Western blotting法检测人食管癌细胞株KYSE140、KYSE150、EC109及EC9706及食管永生化细胞NE3中KLF4蛋白的表达,化学合成2对靶向KLF4的siRNA(KLF4-siRNA1,KLF4-siRNA2),并设对照siRNA(Ctrl-siRNA),分别体外转染至高表达KLF4的食管癌KYSE140细胞中,形成KLF4-siRNA1-KYSE140、KLF4-siRNA2-KYSE140及Ctrl-siRNA-KYSE140细胞,通过MTT实验、Transwell实验分别检测转染后食管癌KYSE140细胞的增殖及迁移。结果:食管癌细胞株KYSE140中KLF4蛋白的表达明显高于KYSE150、EC109及EC9706细胞株[(5.62±0.02)vs(1.71±0.23)、(3.24±0.35)、(3.16±0.41),均P<0.05]。KLF4-siRNA1-KYSE140、KLF4-siRNA2-KYSE140与Ctrl-siRNA-KYSE140细胞相比,KLF4蛋白表达明显降低[(0.49±0.18)、(0.32±0.09)vs(0.98±0.19),均P<0.05],细胞增殖能力明显增高[(1.2±0.8)、(1.4±0.1)vs(0.6±0.1),均P<0.05],迁移细胞数量也明显增加[(780±22)、(475±25)vs(83±17)个,P<0.05]。结论:KLF4在人食管癌细胞的增殖和迁移过程中起着负调控作用。
文摘Salidroside is extensively used as a herbal medicine worldwide, and it has been shown to protect against disruption of endothelial homeostasis and act as an anti-aging agent. The present study aimed to investigate the ameliorative effects of salidroside on homocysteine (Hcy)-induced cell senescence in human umbilical vein endothelial cells (HUVECs) that were mediated via inhibition of Krüppel-like factor 4 (KLF4). An endothelial cell senescence model was induced by Hcy. The cell viability, activities of telomerase and lactate dehydrogenase (LDH), and the level of reactive oxygen species were determined using commercial kits. The expression levels of KLF4, p53 and p21 were determined via western blot analysis, whereas the mRNA expression levels of KLF4 were detected by reverse transcription-quantitative PCR. Small interfering RNA-mediated knockdown of KLF4 was found to reverse Hcy-induced cell senescence. Hcy treatment led to an accelerated cell senescence, as evidenced by decreases in both cell viability and telomerase activity, whereas increases were noted in the leakage of LDH and the level of reactive oxygen species, in addition to an up-regulation of the protein levels of p53 and p21, and up-regulation of KLF4 at both the mRNA and protein level. Treatment with salidroside ameliorated Hcy-induced cell senescence in a dose-dependent manner. Taken together, these results suggested that Hcy may induce cell senescence through upregulation of KLF4, and this may be reversed by treatment with salidroside. Therefore, salidroside was shown to inhibit Hcy-induced cell senescence through KLF4 inhibition.
基金supported by the Start-up Funding of Henan University of Science and Technology(13480027) to Q. C.the Key Science Foundation of Nanjing Medical University(2015NJMUZD002)+2 种基金the Natural Science Foundation of Higher Education Institutions of Jiangsu Province(16KJB-180020)Natural Science Foundation of Jiangsu Province (BK20171053)National Natural Science Funds of China (81702747) to C.L
文摘Kruppel-like factor 4（Klf4） is a zinc finger transcription factor and plays crucial roles in Xenopus embryogenesis.However, its regulation during embryogenesis is still unclear. Here, we report that Tcf711, a key downstream transducer of the Wnt signaling pathway, could promote Klf4 transcription and stimulate Klf4 promoter activity in early Xenopus embryos. Furthermore, cycloheximide treatment showed a direct effect on Klf4 transcription facilitated by Tcf711. Moreover, the dominant negative form of Tcf711（dnTcf711）, which lacks N-terminus of the β-catenin binding motif, could still activate Klf4 transcription, suggesting that this regulation is Wnt/β-catenin independent.Taken together, our results demonstrate that Tcf711 lies upstream of Klf4 to maintain its expression level during Xenopus embryogenesis.
