Objective Hepatic stellate cells(HSCs)play a crucial role in liver fibrosis.Early-stage liver fibrosis is reversible and intimately associated with the state of HSCs.Kruppel-like factor 4(KLF4)plays a pivotal role in ...Objective Hepatic stellate cells(HSCs)play a crucial role in liver fibrosis.Early-stage liver fibrosis is reversible and intimately associated with the state of HSCs.Kruppel-like factor 4(KLF4)plays a pivotal role in a wide array of physiological and pathological processes.This study aimed to investigate the effect of KLF4 on the proliferation,apoptosis and phenotype of quiescent HSCs Methods We designed a KLF4 lentiviral vector and a KLF4 siRNA lentiviral vector,to upregulate and silence KLF4 expression in human HSC LX-2 cells via transfection.Cell proliferation was assessed using the CCK-8 assay.Flow cytometry was used to detect the cell cycle distribution and apoptosis rate.Western blotting was used to determine the levels of some quiescence and activation markers of HSCs Results Overexpression of KLF4 significantly increased the levels of E-cadherin and ZO-1,which are quiescent HSC markers,while significantly decreased the levels of N-cadherin and a-SMA,known activated HSC markers.In contrast,cell proliferation and apoptosis rates were elevated in LX-2 cells in which KLF4 expression was silenced Conclusion KLF4 inhibits the proliferation and activation of human LX-2 HSCs.It might be a key regulatory protein in the maintenance of HSC quiescence and may serve as a target for the inhibition of hepatic fibrosis.展开更多
Although titanate nanofibers(TiNFs)and titanate nanotubes(TiNTs)have been proposed as relatively biocompatible nanomaterials(NMs),there is currently lacking of systemic studies which investigated the toxicity of TiNFs...Although titanate nanofibers(TiNFs)and titanate nanotubes(TiNTs)have been proposed as relatively biocompatible nanomaterials(NMs),there is currently lacking of systemic studies which investigated the toxicity of TiNFs and TiNTs to endothelium.In this study,we developed endothelial monolayer model by using cell culture inserts,and systemically investigated the toxicity of TiNFs and TiNTs by RNA-seq,with a focus on Kruppel-like factor(KLF)-mediated effects,since KLF are transcription factors(TF)involved in the regulation of vascular biology.It was shown that NMs did not significantly induce cytotoxicity despite substantial internalization.However,the expression of many KLF was altered,and Western blot further confirmed that NMs down-regulated KLF2 proteins.Ingenuity pathway analysis(IPA)revealed that NMs altered the expression of KLF2-targed genes,typically the genes involved in inflammatory responses.KLF2-related Gene Ontology(GO)terms and Kyoto Encyclopedia of Gene and Genomes(KEGG)pathways were also altered,and it should be noticed that NMs altered GO terms and KEGG pathways related with endothelial NO synthase(eNOS).This study further verified that NMs decreased intracellular NO and eNOS proteins.All the observed effects were more obvious for TiNFs compared with TiNTs.Combined,this study showed that TiNFs or TiNTs we re non-cytotoxic to endothelial monolayer model,but TiNFs and more modestly TiNTs decreased KLF2 leading to decreased eNOS proteins and NO production.Our data may provide novel understanding about the toxicity of TiNFs as well as other Ti-based NMs to endothelium.展开更多
Objective: To study the correlation of KLF4 and UBE2C expression levels in neuroblastoma with cell adhesion and migration. Methods: A total of 56 children who were diagnosed with neuroblastoma in the Central Hospital ...Objective: To study the correlation of KLF4 and UBE2C expression levels in neuroblastoma with cell adhesion and migration. Methods: A total of 56 children who were diagnosed with neuroblastoma in the Central Hospital of Enshi Autonomous Prefecture between May 2014 and February 2017 were selected as the NB group of the study, and the lesion tissue was collected;38 children who were treated in the Central Hospital of Enshi Autonomous Prefecture due to serious hydronephrosis during the same period were selected as the control group of the study, and the normal adrenal gland tissue was collected. The mRNA expression and protein expression of KLF4 and UBE2C as well as the protein expression of cell adhesion molecules and migration molecules in clinical tissue samples were determined. Results: The mRNA expression and protein expression of KLF4 in neuroblastoma tissue of NB group were greatly lower than those of control group whereas the mRNA expression and protein expression of UBE2C were greatly higher than those of control group;PDLIM1, AMF, GPx1, L1CAM, Nrg1, RANK, RANKL, Inβ1, MTA1 and MMP9 protein expression in neuroblastoma tissue of NB group were greatly higher than those of control group, negatively correlated with the protein expression KLF4, and positively correlated with the protein expression of UBE2C. Conclusion: The low expression of KLF4 and the high expression of UBE2C in neuroblastoma can promote the adhesion and migration of tumor cells.展开更多
基金supported by the National Natural Science Foundation of China(No.81071541).
文摘Objective Hepatic stellate cells(HSCs)play a crucial role in liver fibrosis.Early-stage liver fibrosis is reversible and intimately associated with the state of HSCs.Kruppel-like factor 4(KLF4)plays a pivotal role in a wide array of physiological and pathological processes.This study aimed to investigate the effect of KLF4 on the proliferation,apoptosis and phenotype of quiescent HSCs Methods We designed a KLF4 lentiviral vector and a KLF4 siRNA lentiviral vector,to upregulate and silence KLF4 expression in human HSC LX-2 cells via transfection.Cell proliferation was assessed using the CCK-8 assay.Flow cytometry was used to detect the cell cycle distribution and apoptosis rate.Western blotting was used to determine the levels of some quiescence and activation markers of HSCs Results Overexpression of KLF4 significantly increased the levels of E-cadherin and ZO-1,which are quiescent HSC markers,while significantly decreased the levels of N-cadherin and a-SMA,known activated HSC markers.In contrast,cell proliferation and apoptosis rates were elevated in LX-2 cells in which KLF4 expression was silenced Conclusion KLF4 inhibits the proliferation and activation of human LX-2 HSCs.It might be a key regulatory protein in the maintenance of HSC quiescence and may serve as a target for the inhibition of hepatic fibrosis.
基金financially supported by Hunan Innovative Province Construction Special Major Landmark Innovation Demonstration Project(No.2019XK2303)Xiangtan Science and Technology Project(No.ZD-ZD20191007)。
文摘Although titanate nanofibers(TiNFs)and titanate nanotubes(TiNTs)have been proposed as relatively biocompatible nanomaterials(NMs),there is currently lacking of systemic studies which investigated the toxicity of TiNFs and TiNTs to endothelium.In this study,we developed endothelial monolayer model by using cell culture inserts,and systemically investigated the toxicity of TiNFs and TiNTs by RNA-seq,with a focus on Kruppel-like factor(KLF)-mediated effects,since KLF are transcription factors(TF)involved in the regulation of vascular biology.It was shown that NMs did not significantly induce cytotoxicity despite substantial internalization.However,the expression of many KLF was altered,and Western blot further confirmed that NMs down-regulated KLF2 proteins.Ingenuity pathway analysis(IPA)revealed that NMs altered the expression of KLF2-targed genes,typically the genes involved in inflammatory responses.KLF2-related Gene Ontology(GO)terms and Kyoto Encyclopedia of Gene and Genomes(KEGG)pathways were also altered,and it should be noticed that NMs altered GO terms and KEGG pathways related with endothelial NO synthase(eNOS).This study further verified that NMs decreased intracellular NO and eNOS proteins.All the observed effects were more obvious for TiNFs compared with TiNTs.Combined,this study showed that TiNFs or TiNTs we re non-cytotoxic to endothelial monolayer model,but TiNFs and more modestly TiNTs decreased KLF2 leading to decreased eNOS proteins and NO production.Our data may provide novel understanding about the toxicity of TiNFs as well as other Ti-based NMs to endothelium.
文摘Objective: To study the correlation of KLF4 and UBE2C expression levels in neuroblastoma with cell adhesion and migration. Methods: A total of 56 children who were diagnosed with neuroblastoma in the Central Hospital of Enshi Autonomous Prefecture between May 2014 and February 2017 were selected as the NB group of the study, and the lesion tissue was collected;38 children who were treated in the Central Hospital of Enshi Autonomous Prefecture due to serious hydronephrosis during the same period were selected as the control group of the study, and the normal adrenal gland tissue was collected. The mRNA expression and protein expression of KLF4 and UBE2C as well as the protein expression of cell adhesion molecules and migration molecules in clinical tissue samples were determined. Results: The mRNA expression and protein expression of KLF4 in neuroblastoma tissue of NB group were greatly lower than those of control group whereas the mRNA expression and protein expression of UBE2C were greatly higher than those of control group;PDLIM1, AMF, GPx1, L1CAM, Nrg1, RANK, RANKL, Inβ1, MTA1 and MMP9 protein expression in neuroblastoma tissue of NB group were greatly higher than those of control group, negatively correlated with the protein expression KLF4, and positively correlated with the protein expression of UBE2C. Conclusion: The low expression of KLF4 and the high expression of UBE2C in neuroblastoma can promote the adhesion and migration of tumor cells.
