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connective tissue growth factor expression hints at aggressive nature of colorectal cancer

World Journal of Gastroenterology 2022年第5期28卷 547-569页

作者： Ishrat Parveiz Bhat Tahseen Bilal Rather Irfan Maqbool Gowhar Rashid Kulsum Akhtar Gulzar A Bhat Fazl Q Parray Besina Syed Ishrat Younas Khan Mohsin Kazi Muhammad D Hussain Mudassar Syed Department of Clinical Biochemistry Sher-I-Kashmir Institute of Medical SciencesSrinagar 190011Jammu and KashmirIndia Department of General Surgery Sher-I-Kashmir Institute of Medical SciencesSrinagar 190011Jammu and KashmirIndia Department of Pathology Sher-I-Kashmir Institute of Medical SciencesSrinagar 190011Jammu and KashmirIndia Department of Pharmaceutics College of PharmacyKing Saud UniversityPO Box 2457Riyadh 11451Saudi Arabia Department of Pharmaceutical and Biomedical Sciences California Health Sciences UniversityCaliforniaCA 93612United States

BACKGROUND connective tissue growth factor(CTGF)is a mediator of transforming growth factor-beta signaling and plays a key role in connective tissue remodeling,inflammatory processes and fibrosis in various illnesses including *** To investigate the role of CTGF in colorectal cancer(CRC)progression and to compare the CTGF expression with different clinicopathological *** Real-time polymerase chain reaction,immunohistochemistry and Western blotting was performed to evaluate the CTGF expression and the results were statistically analyzed against the clinicopathological variables of patient data using STATA software version *** CTGF expression levels in tumor specimens were significantly higher than their paired normal *** higher protein expression levels showed a significant association with smoking,staging,tumor grade,invasion depth,necrosis of tumor tissue,and both lymphovascular and perineural *** per the cox regression model and classification tree analysis,tumor-nodemetastasis stage and perineural invasion were important predictors for CTGF expression and prognosis of CRC *** analysis indicated that CTGF overexpression was associated with poorer overall and disease-free *** Expression of CTGF was increased in CRC and was linked with poor overall and disease-free survival of CRC *** findings support prior observations and thus CTGF may be a possible prognostic marker in CRC.

关键词： connective tissue growth factor Quantitative real time polymerase chain reaction Immunohistochemistry Western blotting Colorectal cancer

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connective tissue growth factor Expression in Rabbit Corneal Fibroblast and Its Effect on Fibroblast Proliferation In Vitro

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2003年第1期23卷 81-84页

作者：黄琼胡燕华李琦涵王炯 DepartmentofOphthalmology XieheHospitalTongjiMedicalCollegeHuazhongUniversityofScienceandTechnology InstituteofMedicalBiology ChineseAcademyofMedicalSciencesandPekingUnionMedicalCollege

The connective tissue growth factor (CTGF) expression in cultured corneal fibroblasts and its effect on corneal fibroblasts proliferation in vitro were examined. Total RNA was extracted from early passaged rabbit corneal fibroblasts by guanidine isothiocyanate one-step method and mRNA was reversely transcripted into complementary DNA (cDNA). Specific CTGF primers were used in the PCR reaction and the products were analyzed by electrophoresis to determine the expression of mRNA for CTGF against DNA marker. House-keeping gene GAPDH was used as control. Different concentrations of CTGF (0.5, 5, 50, 500, 5 000, 50 000 ng/ml) were added into the third passaged corneal fibroblast culture system, and its effect on corneal fibroblast proliferation was measured by MTT method. The results showed that compared with a GAPDH 450 bp band, CTGF RT-PCR product showed a specific 120 bp band as expected. CTGF produced a dose-dependent increase in the proliferation of corneal fibroblasts but it inhibited fibroblast proliferation at higher concentrations (5 000 and 50 000 ng/ml). It was concluded that proliferating corneal fibroblasts produce CTGF and CTGF helps to promote corneal fibroblast proliferation. The results indicated that CTGF might be involved in the corneal wound healing after photorefractive keratectomy in which corneal fibroblasts are activated to proliferate and secrete growth factors that in turn promote corneal fibroblast proliferation.

关键词： connective tissue growth factor corneal fibroblasts expression proliferation rabbit

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connective tissue growth factor hammerhead ribozyme attenuates human hepatic stellate cell function

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World Journal of Gastroenterology 2009年第30期15卷 3807-3813页

作者： Run-Ping Gao David R Brigstock The Research Institute of Liver Diseases First Hospital Jilin University Changchun 130021 Jilin Province China The Research Institute at Nationwide Children's Hospital Columbus OH United States Division of Pediatric Surgery Department of Surgery The Ohio State University Columbus OH 43205 United States

AIM： To determine the effect of hammerhead ribozyme targeting connective tissue growth factor （CCN2） on human hepatic stellate cell （HSC） function.METHODS： CCN2 hammerhead ribozyme cDNA plus two self-cleaving sequences were inserted into pTriEx2 to produce pTriCCN2-Rz. Each vector was individually transfected into cultured LX-2 human HSCs, which were then stimulated by addition of transforming growth factor （TGF）-β1 to the culture medium. Semiquantitative RT-PCR was used to determine mRNA levels for CCN2 or collagen I, while protein levels of each molecule in cell /ysates and conditioned medium were measured by ELISA. Cell-cycle progression of the transfected cells was assessed by flow cytometry.RESULTS： In pTriEx2-transfected LX-2 cells, TGF-β1 treatment caused an increase in the mRNA level for CCN2 or collagen I, and an increase in produced and secreted CCN2 or extracellular collagen I protein levels, pTriCCN2-Rz-transfected LX-2 cells showed decreased basal CCN2 or collagen mRNA levels, as well as produced and secreted CCN2 or collagen I protein. Furthermore, the TGF-β1-induced increase in mRNA or protein for CCN2 or collagen I was inhibited partially in pTriCCN2-Rz-transfected LX-2 cells. Inhibition of CCN2 using hammerhead ribozyme cDNA resulted in fewer of the cells transitioning into S phase.CONCLUSION： Endogenous CCN2 is a mediator of basal or TGF-β1-induced collagen I production in human HSCs and regulates entry of the cells into S phase.

关键词： connective tissue growth factor Fibrosis Hepatic stellate cell Transforming growth factor-β1

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connective tissue growth factor is associated with the early renal hypertrophy in uninephrectomized diabetic rats

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Chinese Medical Journal 2006年第12期119卷 1010-1016页

作者： LIU Bi-cheng HUANG Hai-quan LUO Dong-dong MA Kun-ling LIU Dian-ge LIU Hong Institute of Nephrology Zhong Da Hospital Southeast UniversityNanjing 210009 China

Background Renal hypertrophy has been regarded as the early feature of diabetic nephropathy （DN）, which may eventually lead to proteinuria and renal fibrosis. However, the exact mechanism of renal hypertrophy is still unclear. The aim of this study was to investigate the possible association of connective tissue growth factor （CTGF） with renal hypertrophy in uninephrectomized diabetic rats. Methods Seventy-two Sprague-Dawley （SD） rats were randomly divided into two groups： control group （group C, n=32） and diabetic nephropathy （group DN, n=40）. Each group was re-divided into 4 subgroups according to the experimental period. The rats were sacrificed at 1, 2, 4, and 8 weeks respectively after induction of diabetes. Diabetes was induced by intraperitoneal injection of streptozotocin （STZ） after rats had received uninephrectomy. Blood glucose （BG）, body weight （BW）, 24-h urinary albumin excretion （24hUalb）, kidney weight （KW）, KW/BW, glomerular tuft area （AG）, glomerular tuft volume （VG）, proximal tubular area （AT） at each time point, the width of glomerular basement membrane （GBM） and tubular basement membrane （TBM） at week 8 were measured when the rats were sacrificed. Renal expression of CTGF and p27kipl were detected by immnnohistochemical staining. The relationship between CTGF expression and increasing of VG and AT was analyzed. Results There was a significant increase of 24hUalb, KW, and KW/BW from week 1 onward in diabetic rats compared to those in group C （P〈0.05, respectively）, diabetic rats also had a significant increase of AG, VG, and AT from week 1 onward. It was also shown that diabetic rats had a thickening of GBM [（245.7±103.0） nm vs （121.8±19.1） nm, P〈0.01] and TBM [（767.7±331.1） nm vs （293.0±110.5） nm, P〈0.01] at week 8. There was a weak expression for CTGF and p27kipl in normal glomeruli and tubuli, while a significant increasing expression of CTGF and p27kipl was found in glomeruli and tubuli in diabetic kidney from week 1 onward （P〈0.05, respec

关键词： connective tissue growth factor p27kip1 diabetic nephropathies hypertrophy

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connective tissue growth factor Expression in Human Bronchial Epithelial Cells

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Acta Biochimica et Biophysica Sinica 2006年第1期38卷 53-57页

作者： AmritaDOSANJH Department of Molecular and Experimental Medicine the Scripps Research Institute USA La Jolla 92122

connective tissue growth factor (CTGF) is a cysteine-rich protein that promotes extracellularmatrix *** is selectively induced by transforming growth factor β and des-Arg kallidin in lungfibroblasts and increases steady-state mRNA levels of α type I collagen,5α-integrin and fibronectin *** epithelial cells have been proposed to functionally interact with lung *** investigated if bronchial epithelial cells are able to synthesize *** bronchial epithelialcells were grown to subconfluence in standard growth *** cells grown in small airwaygrowth media were harvested following starvation for up to 24 *** of CTGF transcripts wasmeasured by *** was also completed using a commercially available *** cells expressed readily detectable CTGF *** of these cells resulted in a quantitativedecline of CTGF *** sequencing of the PCR product identified human ***-tochemistry confirmed intracellular CTGF in the cells and none in negative control *** conclude thatbronchial epithelial cells could be a novel source of *** epithelial cell-derived CTGF could thusdirectly influence the deposition of collagen in certain fibrotic lung diseases.

关键词： bronchial epithelial cell connective tissue growth factor collagen deposition remodeling

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connective tissue growth factor reacts as an IL-6/STAT3-regulated hepatic negative acute phase protein

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World Journal of Gastroenterology 2011年第2期17卷 151-163页

作者： Olav A Gressner Ieva Peredniene Axel M Gressner Institute of Clinical Chemistry and Pathobiochemistry Central LaboratoryRWTH-University Hospital Wisplinghoff Medical Laboratories Classen-Kappelmann-Str.2450931 CologneGermany Synlab Lab Services Gubener Str.3986156 AugsburgGermany

AIM:To investigate the mechanisms involved in a possible modulator role of interleukin(IL) -6 signalling on CYR61-CTGF-NOV(CCN) 2/connective tissue growth factor(CTGF) expression in hepatocytes(PC) and to look for a relation between serum concentrations of these two parameters in patients with acute inflammation. METHODS:Expression of CCN2/CTGF,p-STAT3,p-Smad 3/1 and p-Smad2 was examined in primary freshly isolated rat or cryo-preserved human PC exposed to various stimuli by Western blotting,electrophoretic mobility shift assay(EMSA) ,reporter-gene-assays and reversetranscriptase polymerase chain reaction. RESULTS:IL-6 strongly down-regulated CCN2/CTGF protein and mRNA expression in PC,enhanceable by extracellular presence of the soluble IL-6 receptor gp80,and supported by an inverse relation between IL-6 and CCN2/CTGF concentrations in patients'*** inhi-bition of TGFβ1 driven CCN2/CTGF expression by IL-6 did not involve a modulation of Smad2(and Smad1/3) ***,the STAT3 SH2 domain binding peptide,a selective inhibitor of STAT3 DNA binding activity,counteracted the inhibitory effect of IL-6 on CCN2/CTGF expression much more pronounced than pyrrolidine-dithiocarbamate,an inhibitor primarily of STAT3 *** EMSA confirmed STAT3 binding to the proposed proximal STAT binding site in the CCN2/CTGF promoter. CONCLUSION:CCN2/CTGF is identified as a hepatocellular negative acute phase protein which is downregulated by IL-6 via the STAT3 pathway through interaction on the DNA binding level.

关键词： Hepatocytes Interleukin-6 connective tissue growth factor STAT3 Liver fibrosis Acute phase reaction

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connective tissue growth factor is overexpressed in human hepatocellular carcinoma and promotes cell invasion and growth

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World Journal of Gastroenterology 2012年第47期18卷 7070-7078页

作者： Ming Xiu Ya-Hui Liu David R Brigstock Fang-Hui He Rui-Juan Zhang Run-Ping Gao Department of Hepatic-Biliary-Pancreatic Medicine First HospitalJilin University Department of Hepatic-Biliary-Pancreatic Surgery First HospitalJilin University the Research Institute at Nationwide Children's Hospital ColumbusOH 43205United States Division of Pediatric Surgery Department of Surgerythe Ohio State UniversityColumbusOH 43205United States

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 ***: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 *** image analysis was performed to measure integrated optimal density of CCN2 mRNA-positive cells in carcinoma foci and the surrounding ***-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 ***2 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 ***:In situ hybridization analysis showed that TGF-β1 mRNA was mainly detected in connective tissues and vasculature around carcinoma *** 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 ***-mesenchymal transition phenotype related with CCN2 was detected in 12/36(33.3%) of HCC liver samples at the edges between carcinoma foci and *** 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) ***-β1-induced cell invasion was abrogated by a neutralizing CCN2 antibody showing that CCN2 is a downstream mediator o

关键词： connective tissue growth factor Hepatocellular carcinoma Hepatoma cell line Migration Invasion

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Full composition granules of Huanglian (Rhizoma Coptidis) decrease the serum monocyte chemotactic protein-1 and connective tissue growth factor levels and inhibit kidney nuclear factor-κB expression in rats with high-fat diet-induced diabetes

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Journal of Traditional Chinese Medicine 2021年第3期41卷 424-431页

作者： GUO Yangzhi DU Juan JIANG Min GUO Wei Department of Traditional Chinese Medicine Capital Medical University Affiliated Beijing Shijitan HospitalBeijing 100038China Department of Obstetrics and Gynecology Beijing Changping Hospital of Integrated Chinese and Western MedicineBeijing 102208China Department of Traditional Chinese Medicine the Mongolian Medical and Traditional Chinese Medicine Hospital of WuhaiWuhai 016000China

OBJECTIVE: To investigate the efficacy of the full composition granules of Huanglian(Rhizoma Coptidis)(FGC) on the serum monocyte chemotactic protein-1(MCP-1) and connective tissue growth factor(CTGF) levels and kidney nuclear factor-κB(NF-κB) expression in rats with high-fat diet-induced ***: Diabetes was induced in rats by feeding a high-fat chow combined with intravenous streptozotocin injection. Forty diabetic SpragueDawley rats were randomly assigned to a normal group(NG), model group(MG), irbesartan group(IG), and low-, middle-, and high-dosage FGC groups(LFGC, MFGC, HFGC), with eight rats per group. The IG rats received 31.25 mg·kg^(-1)·d^(-1) irbesartan tablets, whereas those in the LFGC, MFGC,and HFGC were administered 52, 312.5, and 625 mg·kg^(-1)·d^(-1) FGC, respectively. After 12 weeks,bodyweight(BW), left kidney weight(KW), hemoglobin A1c(HbA1c), serum creatinine(Scre), blood urea nitrogen(BUN), and serum MCP-1 and CTGF levels were determined, pathological changes of the kidney were recorded, and kidney NF-κB p65(A) expression was ***: The 24-h urine albumin and levels of HbA1c, Scre, BUN, and serum MCP-1 and CTGF were significantly increased in in the MG compared with the NG, as was the kidney NF-κB(p65) expression(P < 0.05). Furthermore, clear pathological changes in kidney fibrosis were observed in the MG rats. Following irbesartan and FGC administration,the 24-h urine albumin and the levels of HbA1c,Scre, and serum MCP-1 and CTGF were significantly decreased in FCG groups compared with those in the MG, which is in agreement with the change in the kidney NF-κB(p65) expression, whereas the similarly significant decrease only exist in 24-h urine albumin and the levels of serum CTGF after irbesartan administration. Hematoxylin-eosin(HE)staining results indicated that the fibrosis observed in the MG samples was alleviated through FGC ***: FGC may alleviate potential kidney injury by decreasing the serum MCP-1 an

关键词： Huanglian(Rhizoma Coptidis) diabetic nephropathies chemokine CCL2 connective tissue growth factor NF-kappa B

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Effect of transforming growth factor-beta on activity of connective tissue growth factor gene promoter in mouse NIH/3T3 fibroblasts

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Acta Pharmacologica Sinica 2004年第4期25卷 87-91页

作者： Qing ZHAO, Nan CHEN2, Wei-ming WANG, Jian LU3,Bing-bing DAI3 Department of Nephrology, Ruijin Hospital 3Department of Biochemistry, Shanghai Second Medical University, Shanghai 200025, China Department of Nephrology Ruijin Hospital

AIM: To investigate the regulatory mechanism of transforming growth factor-betaon activity of connective tissue growth factor promoter in mouse NIH/3T3 fibroblasts. METHODS: The regulation fragment of the 5' flanking region of the human CTGF gene was linked to pGL3-Basic vector, a firefly luciferase reporter construct without promoter. The recombinant plasmid pCTGF-luc was transiently transfected to NIH/3T3 fibroblasts. The activity of CTGF promoter after treatment of TGF-β1 and MAPK pathway inhibitors were assayed with luciferase reporter gene assay system. RESULTS: TGF-β1-induced increase of CTGF promoter activity was concentration-dependent, with a plateau at 5 μg/L by 2.67-fold vs control (P<0.05). The TGF-β1 stimulation of CTGF promoter activity was time-dependent, too. After exposure to TGF-β1(5 μg/L), the maximal level of luciferase activity was reached at 12 h and maintained to 24 h by 2.76- and 2.20-fold vs control, respectively (P<0.05). Blockade of mitogen-activated protein kinases (MAPK) pathway with PD98059 (10 μmol/L), the MAP kinase kinase 1 inhibitor, and SB203580 (10 μmol/L), the p38 MAP kinaseinhibitor, decreased basal and TGF-β1-induced activation of CTGF promoter. However, inhibition of c-Jun-N-terminal kinase/stress-activated protein kinase by SP600125 (20 μmol/L) was without effect. CONCLUSION: TGF-β1stimulated the transcriptional activity of CTGF gene promoter in NIH/3T3 fibroblasts in a dose- and time-dependent manner. MAPK pathway may play a role in the regulation of TGF-β1-induced CTGF expression.

关键词： transforming growth factor-beta connective tissue growth factor promoter mitogen-activated protein kinases signal transduction

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Molecular evolution of connective tissue growth factor in Cyprinidae (Teleostei:Cypriniformes)

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Progress in Natural Science:Materials International 2008年第2期18卷 155-160页

作者： Xianghui Kong a,b,Xuzhen Wang a,Xiaoni Gan a,Junbing Li a,Shunping He a,a Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan 430072,China b College of Life Science,Henan Normal University,Xinxiang 453007,China Institute of Hydrobiology Chinese Academy of Sciences Wuhan 430072 China College of Life Science Henan Normal University Xinxiang 453007 China

connective tissue growth factor (CTGF) plays an important role in regulation of cell growth,differentiation,apoptosis and individual development in animals. The study of sequences variation and molecular evolution of CTGF gene across various species of the cyprinid could be helpful for understanding of speciation and gene divergence in this kind of fish. In this study,19 novel sequences of CTGF gene were obtained from the representative species of the family Cyprinidae using PCR amplification,cloning and sequencing. Phylogenetic relationships of Cyprinidae were reconstructed by neighbor-joining (NJ),maximum parsimony (MP),maximum likelihood (ML) and Bayesian method. Oryzias latipes from the family Cyprinodontidae was assigned to be the outgroup taxon. Leuciscini and Barbini were clustered into the monophyletic lineages,respectively,with the high nodal supports. The estimation of the ratio of non-synonymous to synonymous substitution (dN/dS) for the various branches indicated that there stood the different evolution rates between the Leuciscini and the Barbini. With the ratio of dN/dS of the Leuciscini being lower than that of the Barbini,species within the Barbini were demonstrated to be subjected to the relatively less selection pressure and under the relaxable evolution background. A 6 bp indel (insertion/deletion) was found at the 50 end of CTGF gene of Cyprinidae,and this 6 bp deletion only appeared in the Leuciscini,which is a typical characteristic of the Leuciscini and provides evidence for the monophylogeny of the Leuciscini. For the amino acid sequences of CTGF protein,the most variations and indels were distributed in the signal region and IGFBP region of this protein,implying that these variations were correlated with the regulation of the CTGF gene expression and protein activity.

关键词： connective tissue growth factor Sequence variation Molecular evolution Cyprinidae

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