AIM To determine the association of human antigen R(HuR) and inhibitors of apoptosis proteins(IAP1, IAP2) and prognosis in pancreatic cancer.METHODS Protein and mRNA expression levels of IAP1, IAP2 and HuR in pancreat...AIM To determine the association of human antigen R(HuR) and inhibitors of apoptosis proteins(IAP1, IAP2) and prognosis in pancreatic cancer.METHODS Protein and mRNA expression levels of IAP1, IAP2 and HuR in pancreatic ductal adenocarcinoma(PDAC) were compared with normal pancreatic tissue. The correlations among IAP1/IAP2 and HuR as well as their respective correlations with clinicopathological parameters were analyzed. The Kaplan-Meier method and log-rank tests were used for survival analysis. Immunoprecipitation assay was performed to demonstrate HuR binding to IAP1, IAP2 mRNA. PANC1 cells were transfected with either anti-HuR siRNA or control siRNA for 72 h and quantitative reverse transcription polymerase chain reaction(RT-PCR), western blot analysis was carried out.RESULTS RT-PCR analysis revealed that HuR, IAP1, IAP2 mRNA expression were accordingly 3.3-fold, 5.5-fold and 8.4 higher in the PDAC when compared to normal pancreas(P < 0.05). Expression of IAP1 was positively strongly correlated with HuR expression(P < 0.05, r = 0.783). Western blot analysis confirmed RTPCR results. High IAP1 expression, tumor resection status, T stage, lymph-node metastases, tumor differentiation grade, perineural and lymphatic invasion were identified as significant factors for shorter survival in PDAC patients(P < 0.05).Immunohistological analysis showed that HuR was mainly expressed in the ductal cancer cell's nucleus and less so in cytoplasm. RNA immunoprecipitation analysis confirmed IAP1 and IAP2 post-transcriptional regulation by HuR protein. Following siHuR transfection, IAP1 mRNA and protein levels were decreased, however IAP2 expression levels were increased.CONCLUSION HuR mediated overexpression of IAP1 significantly correlates with poor outcomes and early progression of pancreatic cancer. Further studies are needed to assess the underlying mechanisms.展开更多
OBJECTIVE: To investigate the mechanism and effect of Renshen(Radix Ginseng) polysaccharide on the migration of intestinal epithelial cell line 6(IEC-6), as well as the repair mechanism of Renshen(Radix Ginseng) polys...OBJECTIVE: To investigate the mechanism and effect of Renshen(Radix Ginseng) polysaccharide on the migration of intestinal epithelial cell line 6(IEC-6), as well as the repair mechanism of Renshen(Radix Ginseng) polysaccharide on colonic injury induced by dextran sulfate sodium(DSS) in mice. METHODS: Mice were fed 3%(w/v) DSS for 6 d to create colonic lesions. A cell-migration model was created using cell scratching. m RNA expression, protein expression, translation efficiency of m RNA, and nucleoplasmic distribution of human antigen R(Hu R) were determined by real-time reverse transcription-quantitative polymerase chain reaction, western blotting, a dual luciferase reporter system, and immunofluorescence staining, respectively. RESULTS: Renshen(Radix Ginseng) polysaccharide promoted the migration of IEC-6 cells and affected expression of stromal interaction molecule 1(STIM1) and cell division cycle 42(Cdc42) at transcriptional and posttranscriptional levels. CONCLUSIONS: Renshen(Radix Ginseng) polysaccharideinduced repair of intestinal mucosal injury may be mediated by increased cell migration via polyaminebased regulatory mechanisms. In vitro and in vivo experiments suggest that Renshen(Radix Ginseng) polysaccharide-induced post-transcriptional regulation of STIM1 and Cdc42 may be related to differences in the regulation of different target genes by Hu R. Taken together, these data provide a reference for further exploration of the protective effect of Renshen(Radix Ginseng) on the intestinal mucosa.展开更多
Background Human antigen R （HuR） is a ubiquitously expressed member of the ELAV family, and has relatively high cytoplasmic abundance in lung tissue regenerating after injury. In this study, we investigated whether ...Background Human antigen R （HuR） is a ubiquitously expressed member of the ELAV family, and has relatively high cytoplasmic abundance in lung tissue regenerating after injury. In this study, we investigated whether mitogen-activated protein kinase （MAPK）-activated protein kinase 2 （MK2） and HuR participate in the tumor necrosis factor （TNF）-induced expression of interleukin-6 （IL-6）. Methods Human pulmonary microvascular endothelial cells were treated with TNF following short interfering RNAmediated knockdown of MK2 or HuR. Cell supernatants were collected to detect the mRNA and protein expression of IL-6 at different time points, The expression and half-life of IL-6 mRNA were then determined in cells that had been treated with actinomycin D. Finally, after knockdown of MK2, the cytoplasmic expression of HuR protein was analyzed using Western blotting. Results MK2 or HuR knockdown decreased both the mRNA and protein expression of IL-6 in TNF-stimulated cells. In MK2 knockdown cells, the half-life of IL-6 mRNA was reduced to 36 minutes, compared with 67 minutes in the control group. In HuR knockdown cells, the half-life of IL-6 mRNA decreased from 62 minutes to 24 minutes. Further analysis revealed that knockdown of MK2 resulted in reduced HuR protein expression in the cytoplasm. Conclusions MK2 regulates the TNF-induced expression of IL-6 by influencing the cytoplasmic levels of HuR.展开更多
文摘AIM To determine the association of human antigen R(HuR) and inhibitors of apoptosis proteins(IAP1, IAP2) and prognosis in pancreatic cancer.METHODS Protein and mRNA expression levels of IAP1, IAP2 and HuR in pancreatic ductal adenocarcinoma(PDAC) were compared with normal pancreatic tissue. The correlations among IAP1/IAP2 and HuR as well as their respective correlations with clinicopathological parameters were analyzed. The Kaplan-Meier method and log-rank tests were used for survival analysis. Immunoprecipitation assay was performed to demonstrate HuR binding to IAP1, IAP2 mRNA. PANC1 cells were transfected with either anti-HuR siRNA or control siRNA for 72 h and quantitative reverse transcription polymerase chain reaction(RT-PCR), western blot analysis was carried out.RESULTS RT-PCR analysis revealed that HuR, IAP1, IAP2 mRNA expression were accordingly 3.3-fold, 5.5-fold and 8.4 higher in the PDAC when compared to normal pancreas(P < 0.05). Expression of IAP1 was positively strongly correlated with HuR expression(P < 0.05, r = 0.783). Western blot analysis confirmed RTPCR results. High IAP1 expression, tumor resection status, T stage, lymph-node metastases, tumor differentiation grade, perineural and lymphatic invasion were identified as significant factors for shorter survival in PDAC patients(P < 0.05).Immunohistological analysis showed that HuR was mainly expressed in the ductal cancer cell's nucleus and less so in cytoplasm. RNA immunoprecipitation analysis confirmed IAP1 and IAP2 post-transcriptional regulation by HuR protein. Following siHuR transfection, IAP1 mRNA and protein levels were decreased, however IAP2 expression levels were increased.CONCLUSION HuR mediated overexpression of IAP1 significantly correlates with poor outcomes and early progression of pancreatic cancer. Further studies are needed to assess the underlying mechanisms.
基金National Natural Science Foundation of China:Study on the Effect of Supplementing Qi and Invigorating the Spleen on the Migration of Small Intestinal Epithelial Cells through the Regulation of Polyamines and Calcium Ions (No. 81673940)First-class Discipline Construction Major Project of Guangzhou University of Chinese Medicine,Guangzhou University of Chinese Medicine Planning (2020) No. 62:based on the Viscous Characteristics of Dampness and Pathogenic Factors in Lingnan,this Paper Studied the Differential Characteristics of the Syndrome of Deficiency of Spleen and Stomach Disease and the Intervention Mechanism of Lingnan Traditional Chinese Medicine for Mucosal Damage Repair。
文摘OBJECTIVE: To investigate the mechanism and effect of Renshen(Radix Ginseng) polysaccharide on the migration of intestinal epithelial cell line 6(IEC-6), as well as the repair mechanism of Renshen(Radix Ginseng) polysaccharide on colonic injury induced by dextran sulfate sodium(DSS) in mice. METHODS: Mice were fed 3%(w/v) DSS for 6 d to create colonic lesions. A cell-migration model was created using cell scratching. m RNA expression, protein expression, translation efficiency of m RNA, and nucleoplasmic distribution of human antigen R(Hu R) were determined by real-time reverse transcription-quantitative polymerase chain reaction, western blotting, a dual luciferase reporter system, and immunofluorescence staining, respectively. RESULTS: Renshen(Radix Ginseng) polysaccharide promoted the migration of IEC-6 cells and affected expression of stromal interaction molecule 1(STIM1) and cell division cycle 42(Cdc42) at transcriptional and posttranscriptional levels. CONCLUSIONS: Renshen(Radix Ginseng) polysaccharideinduced repair of intestinal mucosal injury may be mediated by increased cell migration via polyaminebased regulatory mechanisms. In vitro and in vivo experiments suggest that Renshen(Radix Ginseng) polysaccharide-induced post-transcriptional regulation of STIM1 and Cdc42 may be related to differences in the regulation of different target genes by Hu R. Taken together, these data provide a reference for further exploration of the protective effect of Renshen(Radix Ginseng) on the intestinal mucosa.
基金This study was supported by the National Natural Science Foundation of China （No. 81270138）, the Natural Science Foundation of Jiangsu Province （No. BK2011657 and No. BK20130402）, and the Medical Technology Innovation Foundation of Nanjing Military Command （No. CWS 12J008）.
文摘Background Human antigen R （HuR） is a ubiquitously expressed member of the ELAV family, and has relatively high cytoplasmic abundance in lung tissue regenerating after injury. In this study, we investigated whether mitogen-activated protein kinase （MAPK）-activated protein kinase 2 （MK2） and HuR participate in the tumor necrosis factor （TNF）-induced expression of interleukin-6 （IL-6）. Methods Human pulmonary microvascular endothelial cells were treated with TNF following short interfering RNAmediated knockdown of MK2 or HuR. Cell supernatants were collected to detect the mRNA and protein expression of IL-6 at different time points, The expression and half-life of IL-6 mRNA were then determined in cells that had been treated with actinomycin D. Finally, after knockdown of MK2, the cytoplasmic expression of HuR protein was analyzed using Western blotting. Results MK2 or HuR knockdown decreased both the mRNA and protein expression of IL-6 in TNF-stimulated cells. In MK2 knockdown cells, the half-life of IL-6 mRNA was reduced to 36 minutes, compared with 67 minutes in the control group. In HuR knockdown cells, the half-life of IL-6 mRNA decreased from 62 minutes to 24 minutes. Further analysis revealed that knockdown of MK2 resulted in reduced HuR protein expression in the cytoplasm. Conclusions MK2 regulates the TNF-induced expression of IL-6 by influencing the cytoplasmic levels of HuR.
