AIM:To determine the expression characteristics of connective tissue growth factor(CTGF/CCN2) in human hepatocellular carcinoma(HCC) in histology and to elucidate the roles of CCN2 on hepatoma cell cycle progression a...AIM:To determine the expression characteristics of connective tissue growth factor(CTGF/CCN2) in human hepatocellular carcinoma(HCC) in histology and to elucidate the roles of CCN2 on hepatoma cell cycle progression and metastasis in vitro.METHODS:Liver samples from 36 patients(who underwent hepatic resection for the first HCC between 2006 and 2011) and 6 normal individuals were examined for transforming growth factor β1(TGF-β1) or CCN2 mRNA by in situ hybridization.Computer image analysis was performed to measure integrated optimal density of CCN2 mRNA-positive cells in carcinoma foci and the surrounding stroma.Fibroblast-specific protein-1(FSP-1) and E-cadherin were examined to evaluate the process of epithelial to mesenchymal transition,α-smooth muscle actin and FSP-1 were detected to identify hepatic stellate cells,and CD34 was measured to evaluate the extent of vascularization in liver tissues by immunohistochemical staining.CCN2 was assessed for its stimulation of HepG2 cell migration and invasion using commercial kits while flow cytometry was used to determine CCN2 effects on HepG2 cell-cycle.RESULTS:In situ hybridization analysis showed that TGF-β1 mRNA was mainly detected in connective tissues and vasculature around carcinoma foci.In comparison to normal controls,CCN2 mRNA was enhanced 1.9-fold in carcinoma foci(12.36 ± 6.08 vs 6.42 ± 2.35) or 9.4-fold in the surrounding stroma(60.27 ± 28.71 vs 6.42 ± 2.35),with concomitant expression of CCN2 and TGF-β1 mRNA in those areas.Epithelial-mesenchymal transition phenotype related with CCN2 was detected in 12/36(33.3%) of HCC liver samples at the edges between carcinoma foci and vasculature.Incubation of HepG2 cells with CCN2(100 ng/mL) resulted in more of the cells transitioning into S phase(23.85 ± 2.35 vs 10.94 ± 0.23),and induced a significant migratory(4.0-fold) and invasive(5.7-fold) effect.TGF-β1-induced cell invasion was abrogated by a neutralizing CCN2 antibody showing that CCN2 is a downstream mediator of TGF-β1-induced hepatoma cell invasion.CONCLUSION:These data support a role for CCN2 in the growth and metastasis of HCC and highlight CCN2 as a potential novel therapeutic target.展开更多
BACKGROUND: Pancreatic stellate cells(PSCs) play a critical role in the pathogenesis of pancreatic fibrosis and have emerging functions as progenitor cells,immune cells or intermediaries in pancreatic exocrine secreti...BACKGROUND: Pancreatic stellate cells(PSCs) play a critical role in the pathogenesis of pancreatic fibrosis and have emerging functions as progenitor cells,immune cells or intermediaries in pancreatic exocrine secretion. Increasing evidence has shown that desmin as an exclusive cytoskeleton marker of PSC is only expressed in part of these cells. This study was to establish a desmin-positive PSC cell line and evaluate its actions on pancreatic fibrosis,inflammation and immunity.METHODS: The presence of cytoskeletal proteins,integrin α5β1 or TLR4,was determined by immunocytochemistry while the production of desmin,collagen I,MMP-1,MMP-2,TIMP-2,or CD14 was evaluated by Western blotting. The levels of desmin,collagen I,IL-1 and IL-6 m RNA were determined by real-time quantitative PCR. The secretion of cytokines was detected by ELISA. Cell function was assessed using adhesion,migration,or proliferation assays. RESULTS: A stable activated rat PSC cell line(designated as RP-2) was established by RSV promoter/enhancer-driven SV40 large T antigen expression. RP-2 cells retained typical PSC properties,exhibited a myofibroblast-like phenotype and persistently produced desmin. The cells produced collagen I protein,matrix metalloproteinases and inhibitors thereof. RP-2 cells demonstrated typical PSC functions,including proliferation,adherence,and migration,the latter two of which occurred in response to fibronectin and were mediated byintegrin α5β1. TLR4 and its response genes including proinflammatory cytokines(IL-1,IL-6,TNF-α) and chemotactic cytokines(MCP-1,MIP-1α,Rantes) were produced by RP-2 cells and activated by LPS. LPS-induced IL-1 or IL-6 m RNA expression in this cell line was fully blocked with My D88 inhibitor.CONCLUSION: RP-2 cells provide a novel tool for analyzing the properties and functions of PSCs in the pathogenesis of fibrosis,inflammation and immunity in the pancreas.展开更多
Type 1 autoimmune pancreatitis(AIP)or chronic sclerosing sialadenitis(Küttner’s tumour)is an uncommon disorder that has recently been confirmed as an Ig G4-related disease.Here,we describe a rare case of a53-yea...Type 1 autoimmune pancreatitis(AIP)or chronic sclerosing sialadenitis(Küttner’s tumour)is an uncommon disorder that has recently been confirmed as an Ig G4-related disease.Here,we describe a rare case of a53-year-old male patient who primarily presented with pancreatic body mass,left neck mass and several lumps in his lower lip mimicking pancreatic cancer(PC)and neck metastasis.The patient underwent pancreatic body mass and labial gland lumps resection as well as an ultrasound-guided biopsy of the left neck mass.He was diagnosed with Ig G4-related focal type of AIP(f-AIP)and Küttner’s tumour by immunohistochemistry.The patient responded well to corticosteroid therapy andremains healthy with no signs of recurrence at one year follow-up.The differentiation of f-AIP from PC is very important to avoid unnecessary pancreatic resection.展开更多
A 61-year-old male from Northeast China presented with a 2-mo history of abdominal distension,pruritus and jaundice.Laboratory testing revealed an elevated serum Ig G4 level.A computed tomography scan showed a typical...A 61-year-old male from Northeast China presented with a 2-mo history of abdominal distension,pruritus and jaundice.Laboratory testing revealed an elevated serum Ig G4 level.A computed tomography scan showed a typical feature of autoimmune pancreatitis(AIP) and cholecystocholangitis.Early gastric cancer was incidentally discovered when endoscopic untrasound-guided fine needle aspiration(EUSFNA) of the pancreas was carried out.The patient underwent radical subtotal gastrectomy for gastric cancer combined with cholecystectomy.Helicobacter pylori(H.pylori) and Ig G4-positive plasmacytes were detected in gastric cancer tissue,pancreatic EUSFNA sample and resected gallbladder specimen by immunohistochemistry.The patient was diagnosed with H.pylori-positive Ig G4-related AIP and sclerosing cholecystocholangitis as well as H.pylori-positive gastric cancer.He responded well to steroid therapy and remains healthy with no signs of recurrence at one year follow-up.We speculate that H.pylori might act as a trigger via direct or indirect action in the initiation of onset of gastric cancer and multiorgan Ig G4-related disease.展开更多
基金Supported by National Natural Scientific Foundation,No. 30872236,81070370,to Gao RPNIH 5R01AA016003,to Brigstock DR
文摘AIM:To determine the expression characteristics of connective tissue growth factor(CTGF/CCN2) in human hepatocellular carcinoma(HCC) in histology and to elucidate the roles of CCN2 on hepatoma cell cycle progression and metastasis in vitro.METHODS:Liver samples from 36 patients(who underwent hepatic resection for the first HCC between 2006 and 2011) and 6 normal individuals were examined for transforming growth factor β1(TGF-β1) or CCN2 mRNA by in situ hybridization.Computer image analysis was performed to measure integrated optimal density of CCN2 mRNA-positive cells in carcinoma foci and the surrounding stroma.Fibroblast-specific protein-1(FSP-1) and E-cadherin were examined to evaluate the process of epithelial to mesenchymal transition,α-smooth muscle actin and FSP-1 were detected to identify hepatic stellate cells,and CD34 was measured to evaluate the extent of vascularization in liver tissues by immunohistochemical staining.CCN2 was assessed for its stimulation of HepG2 cell migration and invasion using commercial kits while flow cytometry was used to determine CCN2 effects on HepG2 cell-cycle.RESULTS:In situ hybridization analysis showed that TGF-β1 mRNA was mainly detected in connective tissues and vasculature around carcinoma foci.In comparison to normal controls,CCN2 mRNA was enhanced 1.9-fold in carcinoma foci(12.36 ± 6.08 vs 6.42 ± 2.35) or 9.4-fold in the surrounding stroma(60.27 ± 28.71 vs 6.42 ± 2.35),with concomitant expression of CCN2 and TGF-β1 mRNA in those areas.Epithelial-mesenchymal transition phenotype related with CCN2 was detected in 12/36(33.3%) of HCC liver samples at the edges between carcinoma foci and vasculature.Incubation of HepG2 cells with CCN2(100 ng/mL) resulted in more of the cells transitioning into S phase(23.85 ± 2.35 vs 10.94 ± 0.23),and induced a significant migratory(4.0-fold) and invasive(5.7-fold) effect.TGF-β1-induced cell invasion was abrogated by a neutralizing CCN2 antibody showing that CCN2 is a downstream mediator of TGF-β1-induced hepatoma cell invasion.CONCLUSION:These data support a role for CCN2 in the growth and metastasis of HCC and highlight CCN2 as a potential novel therapeutic target.
基金support by grants from the National Natural Science Foundation of China(81070370 and 81270544)
文摘BACKGROUND: Pancreatic stellate cells(PSCs) play a critical role in the pathogenesis of pancreatic fibrosis and have emerging functions as progenitor cells,immune cells or intermediaries in pancreatic exocrine secretion. Increasing evidence has shown that desmin as an exclusive cytoskeleton marker of PSC is only expressed in part of these cells. This study was to establish a desmin-positive PSC cell line and evaluate its actions on pancreatic fibrosis,inflammation and immunity.METHODS: The presence of cytoskeletal proteins,integrin α5β1 or TLR4,was determined by immunocytochemistry while the production of desmin,collagen I,MMP-1,MMP-2,TIMP-2,or CD14 was evaluated by Western blotting. The levels of desmin,collagen I,IL-1 and IL-6 m RNA were determined by real-time quantitative PCR. The secretion of cytokines was detected by ELISA. Cell function was assessed using adhesion,migration,or proliferation assays. RESULTS: A stable activated rat PSC cell line(designated as RP-2) was established by RSV promoter/enhancer-driven SV40 large T antigen expression. RP-2 cells retained typical PSC properties,exhibited a myofibroblast-like phenotype and persistently produced desmin. The cells produced collagen I protein,matrix metalloproteinases and inhibitors thereof. RP-2 cells demonstrated typical PSC functions,including proliferation,adherence,and migration,the latter two of which occurred in response to fibronectin and were mediated byintegrin α5β1. TLR4 and its response genes including proinflammatory cytokines(IL-1,IL-6,TNF-α) and chemotactic cytokines(MCP-1,MIP-1α,Rantes) were produced by RP-2 cells and activated by LPS. LPS-induced IL-1 or IL-6 m RNA expression in this cell line was fully blocked with My D88 inhibitor.CONCLUSION: RP-2 cells provide a novel tool for analyzing the properties and functions of PSCs in the pathogenesis of fibrosis,inflammation and immunity in the pancreas.
基金Supported by National Natural Scientific Foundation,No.81070370,and No.81270544 to Gao RPNational Institutes of Health,No.5R01AA016003 to Brigstock DR
文摘Type 1 autoimmune pancreatitis(AIP)or chronic sclerosing sialadenitis(Küttner’s tumour)is an uncommon disorder that has recently been confirmed as an Ig G4-related disease.Here,we describe a rare case of a53-year-old male patient who primarily presented with pancreatic body mass,left neck mass and several lumps in his lower lip mimicking pancreatic cancer(PC)and neck metastasis.The patient underwent pancreatic body mass and labial gland lumps resection as well as an ultrasound-guided biopsy of the left neck mass.He was diagnosed with Ig G4-related focal type of AIP(f-AIP)and Küttner’s tumour by immunohistochemistry.The patient responded well to corticosteroid therapy andremains healthy with no signs of recurrence at one year follow-up.The differentiation of f-AIP from PC is very important to avoid unnecessary pancreatic resection.
基金Supported by National Natural Scientific Foundation of China(to Gao RP),No.81070370 and No.81270544
文摘A 61-year-old male from Northeast China presented with a 2-mo history of abdominal distension,pruritus and jaundice.Laboratory testing revealed an elevated serum Ig G4 level.A computed tomography scan showed a typical feature of autoimmune pancreatitis(AIP) and cholecystocholangitis.Early gastric cancer was incidentally discovered when endoscopic untrasound-guided fine needle aspiration(EUSFNA) of the pancreas was carried out.The patient underwent radical subtotal gastrectomy for gastric cancer combined with cholecystectomy.Helicobacter pylori(H.pylori) and Ig G4-positive plasmacytes were detected in gastric cancer tissue,pancreatic EUSFNA sample and resected gallbladder specimen by immunohistochemistry.The patient was diagnosed with H.pylori-positive Ig G4-related AIP and sclerosing cholecystocholangitis as well as H.pylori-positive gastric cancer.He responded well to steroid therapy and remains healthy with no signs of recurrence at one year follow-up.We speculate that H.pylori might act as a trigger via direct or indirect action in the initiation of onset of gastric cancer and multiorgan Ig G4-related disease.
