AIM:To Investigate the effects of transforming growth factorβ2(TGF-β2)and connective tissue growth factor(CTGF)on transdifferentiation of human lens epithelial cells(HLECs)cultured in vitro and synthesis of extracel...AIM:To Investigate the effects of transforming growth factorβ2(TGF-β2)and connective tissue growth factor(CTGF)on transdifferentiation of human lens epithelial cells(HLECs)cultured in vitro and synthesis of extracellular matrix(ECM).METHODS:HLECs were treated with TGF-β2(0,0.5,1.0,5,10μg/L)and CTGF(0,15,30,60,100μg/L)for different times(0,24,48,72h)in vitro and the expression ofα-smooth muscle actin(α-SMA),the main component of the extracellular matrix typeⅠcollagen(Col-1)and fibronectin(Fn)were measured by using real-time polymerase chain reaction(PCR)and western-blot.RESULTS:TGF-β2 and CTGF significantly increased expression ofα-SMA mRNA and protein(P【0.05,P【0.001),Fn mRNA and protein(P【0.001),Col-1 mRNA and protein(P【0.001).TGF-β2 could induce HLECs expression of CTGF mRNA and protein in dosedependent manner(P【0.05,P【0.001).TGF-β2 and CTGF could induce HLECs to expressα-SMA,Fn and Col-1 in time-dependent manner.Each time of TGF-β2and CTGF induced HELCs expression ofα-SMA,Fn,Col-1 mRNA and protein was significant increase compared with control(P【0.05,P【0.001).CONCLUSION:TGF-β2 and CTGF could induce HLECs epithelial mesenchymal transition and ECM synthesis.展开更多
AIM: To explore the roles of microRNA-let7 c(miR-let7 c) and transforming growth factor-β2(TGF-β2) and cellular signaling during epithelial-to-mesenchymal transition(EMT) of retinal pigment epithelial cells. METHODS...AIM: To explore the roles of microRNA-let7 c(miR-let7 c) and transforming growth factor-β2(TGF-β2) and cellular signaling during epithelial-to-mesenchymal transition(EMT) of retinal pigment epithelial cells. METHODS: Retinal pigment epithelial(ARPE-19) cells were cultured with no serum for 12 h, and then with recombinant human TGF-β2 for different lengths of time. ARPE-19 cells were transfected with 1×106 TU/mL miR-let7 c mimcs(miR-let7 cM), miR-let7 c mimcs negative control(miR-let7cMNC) and miR-let7 c inhibitor(miR-let7 cI) using the transfection reagent. The expression of keratin-18, vimentin, N-cadherin, IKB alpha, p65 were detected by Western blot, quantitative polymerase chain reaction and immunofluorescence. RESULTS: The expression of miR-let7c was dramatically reduced and the nuclear factor-kappa B(NF-κB) signaling pathway was activated after induction by TGF-β2(P<0.05). In turn, overexpressed miR-let7 c significantly inhibited TGF-β2-induced EMT(P<0.05). However, miR-let7 c was unable to inhibit TGF-β2-induced EMT when the NF-κB signaling pathway was inhibited by BAY11-7082(P<0.01). CONCLUSION: The miR-let7 c regulates TGF-β2-induced EMT through the NF-κB signaling pathway in ARPE-19 cells.展开更多
Epidemiological studies have demonstrated that fine particulate matter（PM（2.5）） exposure causes airway inflammation, which may lead to lung cancer. The activation of epithelial–mesenchymal transition（EMT） is as...Epidemiological studies have demonstrated that fine particulate matter（PM（2.5）） exposure causes airway inflammation, which may lead to lung cancer. The activation of epithelial–mesenchymal transition（EMT） is assumed to be a crucial step in lung tumor metastasis and development. We assessed the EMT effect of low concentrations（0, 0.1, 1.0, and 5.0 μg/m L）of PM（2.5） organic extract on a human bronchial epithelial cell line（BEAS-2 B）. PM（2.5） samples were collected from three cities（Shanghai, Ningbo, and Nanjing） in the Yangtze River Delta（YRD） region in autumn 2014. BEAS-2 B cells were exposed to the PM（2.5） extract to assess cell viability, invasion ability as well as the relative m RNA and protein expressions of EMT markers. Our findings revealed that BEAS-2 B cells changed from the epithelial to mesenchymal phenotype after exposure. In all groups, PM（2.5） exposure dose-dependently decreased the expression of E-cadherin and increased the expression of Vimentin. The key transcription factors, including ZEB1 and Slug, were significantly up-regulated upon exposure. These results indicated that the PM（2.5） organic extract induced different degrees of EMT progression in BEAS-2 B cells. The cell invasion ability increased in a concentration-dependent manner after 48 hr of treatment with the extract. This study offers a novel insight into the effects of PM（2.5） on EMT and the potential health risks associated with PM（2.5） in the YRD region.展开更多
基金National Natural Science Foundation of China(No.81070721)Inernational Exchange Program of Shaanxi Province,China(No.2012kw-31)
文摘AIM:To Investigate the effects of transforming growth factorβ2(TGF-β2)and connective tissue growth factor(CTGF)on transdifferentiation of human lens epithelial cells(HLECs)cultured in vitro and synthesis of extracellular matrix(ECM).METHODS:HLECs were treated with TGF-β2(0,0.5,1.0,5,10μg/L)and CTGF(0,15,30,60,100μg/L)for different times(0,24,48,72h)in vitro and the expression ofα-smooth muscle actin(α-SMA),the main component of the extracellular matrix typeⅠcollagen(Col-1)and fibronectin(Fn)were measured by using real-time polymerase chain reaction(PCR)and western-blot.RESULTS:TGF-β2 and CTGF significantly increased expression ofα-SMA mRNA and protein(P【0.05,P【0.001),Fn mRNA and protein(P【0.001),Col-1 mRNA and protein(P【0.001).TGF-β2 could induce HLECs expression of CTGF mRNA and protein in dosedependent manner(P【0.05,P【0.001).TGF-β2 and CTGF could induce HLECs to expressα-SMA,Fn and Col-1 in time-dependent manner.Each time of TGF-β2and CTGF induced HELCs expression ofα-SMA,Fn,Col-1 mRNA and protein was significant increase compared with control(P【0.05,P【0.001).CONCLUSION:TGF-β2 and CTGF could induce HLECs epithelial mesenchymal transition and ECM synthesis.
基金Supported by National Natural Science Foundation of China(No.81600754)。
文摘AIM: To explore the roles of microRNA-let7 c(miR-let7 c) and transforming growth factor-β2(TGF-β2) and cellular signaling during epithelial-to-mesenchymal transition(EMT) of retinal pigment epithelial cells. METHODS: Retinal pigment epithelial(ARPE-19) cells were cultured with no serum for 12 h, and then with recombinant human TGF-β2 for different lengths of time. ARPE-19 cells were transfected with 1×106 TU/mL miR-let7 c mimcs(miR-let7 cM), miR-let7 c mimcs negative control(miR-let7cMNC) and miR-let7 c inhibitor(miR-let7 cI) using the transfection reagent. The expression of keratin-18, vimentin, N-cadherin, IKB alpha, p65 were detected by Western blot, quantitative polymerase chain reaction and immunofluorescence. RESULTS: The expression of miR-let7c was dramatically reduced and the nuclear factor-kappa B(NF-κB) signaling pathway was activated after induction by TGF-β2(P<0.05). In turn, overexpressed miR-let7 c significantly inhibited TGF-β2-induced EMT(P<0.05). However, miR-let7 c was unable to inhibit TGF-β2-induced EMT when the NF-κB signaling pathway was inhibited by BAY11-7082(P<0.01). CONCLUSION: The miR-let7 c regulates TGF-β2-induced EMT through the NF-κB signaling pathway in ARPE-19 cells.
基金supported by the National Natural Science Foundation of China (Nos. 41390240, 21477124, 21677140, 21477123 and 21507128)the Knowledge Innovation Program of the Chinese Academy of Sciences (Nos. IUEMS201405,IUEQN201506)+1 种基金the Science and Technology Program of Fujian Province (No. 2016 T3005)the grant from Xiamen Municipal Bureau of Science and Technology Program (No. 3502Z20161236)
文摘Epidemiological studies have demonstrated that fine particulate matter（PM（2.5）） exposure causes airway inflammation, which may lead to lung cancer. The activation of epithelial–mesenchymal transition（EMT） is assumed to be a crucial step in lung tumor metastasis and development. We assessed the EMT effect of low concentrations（0, 0.1, 1.0, and 5.0 μg/m L）of PM（2.5） organic extract on a human bronchial epithelial cell line（BEAS-2 B）. PM（2.5） samples were collected from three cities（Shanghai, Ningbo, and Nanjing） in the Yangtze River Delta（YRD） region in autumn 2014. BEAS-2 B cells were exposed to the PM（2.5） extract to assess cell viability, invasion ability as well as the relative m RNA and protein expressions of EMT markers. Our findings revealed that BEAS-2 B cells changed from the epithelial to mesenchymal phenotype after exposure. In all groups, PM（2.5） exposure dose-dependently decreased the expression of E-cadherin and increased the expression of Vimentin. The key transcription factors, including ZEB1 and Slug, were significantly up-regulated upon exposure. These results indicated that the PM（2.5） organic extract induced different degrees of EMT progression in BEAS-2 B cells. The cell invasion ability increased in a concentration-dependent manner after 48 hr of treatment with the extract. This study offers a novel insight into the effects of PM（2.5） on EMT and the potential health risks associated with PM（2.5） in the YRD region.
