AIM: To examine the role of Fibrinogen-like protein 2 (fgl2)/fibroleukin in tumor development. Fgl2 has been reported to play a vital role in the pathogenesis in MHV-3 (mouse hepatitis virus) induced fulminant an...AIM: To examine the role of Fibrinogen-like protein 2 (fgl2)/fibroleukin in tumor development. Fgl2 has been reported to play a vital role in the pathogenesis in MHV-3 (mouse hepatitis virus) induced fulminant and severe hepatitis, spontaneous abortion, allo- and xenograft rejection by mediating "immune coagulation".METHODS: Tumor tissues from 133 patients with six types of distinct cancers and the animal tumor tissues from human hepatocellular carcinoma (HCC) model on nude mice (established from high metastasis HCC cell line MHCC97LM6) were obtained. RESULTS: HfgI2 was detected in tumor tissues from 127 out of 133 patients as well as tumor tissues collected from human HCC nude mice. Hfgl2 was highly expressed both in cancer cells and interstitial inflammatory cells including macrophages, NK cells, and CD8^+ T lymphocytes and vascular endothelial cells. HfgI2 mRNA was localized in cells that expressed hfgI2 protein. Fibrin (nogen) colocalization with hfgl2 expression was determined by dual immunohistochemical staining. In vitro, IL-2 and IFN-γ, increased hfgl2 mRNA by 10-100 folds and protein expression in both THP-1 and HUVEC cell lines. One-stage clotting assays demonstrated that THP-1 and HUVEC cells expressing hfgl2 had increased procoagulant activity following cytokines stimulation. CONCLUSION: The hfgI2 contributes to the hypercoagulability in cancer and may induce tumor angiogenesis and metastasis via cytokine induction.展开更多
目的:探讨纤维介素蛋白2凝血酶原酶(fibrinogen-like protein 2/fibroleukin,fgl2)在人肝癌(hepatocel-lular carcinoma,HCC)细胞系HCCLM6细胞的表达及在肿瘤生长中的作用。方法:用RT-PCR方法检测HCCLM6细胞在静息和细胞因子作用下fgl2m...目的:探讨纤维介素蛋白2凝血酶原酶(fibrinogen-like protein 2/fibroleukin,fgl2)在人肝癌(hepatocel-lular carcinoma,HCC)细胞系HCCLM6细胞的表达及在肿瘤生长中的作用。方法:用RT-PCR方法检测HCCLM6细胞在静息和细胞因子作用下fgl2mRNA水平的表达;用RNA干扰技术建立fgl2敲除的HCCLM6细胞模型;以流式细胞技术检测fgl2敲除后的HCCLM6细胞体外培养细胞周期和增殖指数的改变以及对TNF-α(肿瘤坏死因子-α)诱导凋亡的抵抗能力改变。结果:IFN-γ(干扰素-γ)、IL-1β(白介素-1β)和TNF-α均可在mRNA水平上调HCCLM6细胞fgl2的表达,以TNF-α诱导作用最为显著;fgl2干扰后HCCLM6细胞增殖指数显著降低,细胞周期分析S期比例显著下降;fgl2干扰对HCCLM6细胞在全培养基和无血清培养基中培养的凋亡率无显著影响,但在合并大剂量TNF-α诱导下,fgl2干扰可显著提高HCCLM6细胞的凋亡率。结论:炎性细胞因子可诱导HCCLM6细胞表达fgl2,fgl2的表达可直接促进肿瘤细胞的生长,并增强对TNF-α诱导凋亡的耐受。展开更多
The aim of this study is to investigate the important regulative elements region which plays an important role on the activation of transcription exerted by the 5' noncoding region of hfgl2 gene in response to HBc...The aim of this study is to investigate the important regulative elements region which plays an important role on the activation of transcription exerted by the 5' noncoding region of hfgl2 gene in response to HBc and HBx. A series of promoter luciferase report plasmids, in which the hfgl2 gene has been deleted of the 5' and retained the common 3', were constructed. All the plasmids constructed were subjected to electrophoretic analysis and DNA sequencing. A eukaryotic construct expressing HBc or HBx, a luciferase reporter construct containing hfgl2 promoter and aβ-galactosidase (β-gal) plasmid were co-transfected into Chinese hamster ovary (CHO) cells and hepG2 cells, respectively. Luciferase report plasmids containing hfgl2 promoter were successfully constructed, and a serial assays of deletion of hfgl2 gene promoter showed that a strong regulatory region from -817 to -467 (relative to the transcription start site) was responsible for transcription and expression regulation of hfgl2 gene. The important regulative elements region in the promoter of hfgl2 gene was in response to HBc and HBx. which contributes to further pursuit of cis-acting elements and transcriptional factors involved in the transcription of hfgl2 gene.展开更多
基金The Natural Science Foundation of China,NSFC (30672380,30571643)National Key Basic Research Program of China (2007CB512900,2005CB522901,2005CB522507)11th Five-Year Plan Key Project (2006BAI05A07)
文摘AIM: To examine the role of Fibrinogen-like protein 2 (fgl2)/fibroleukin in tumor development. Fgl2 has been reported to play a vital role in the pathogenesis in MHV-3 (mouse hepatitis virus) induced fulminant and severe hepatitis, spontaneous abortion, allo- and xenograft rejection by mediating "immune coagulation".METHODS: Tumor tissues from 133 patients with six types of distinct cancers and the animal tumor tissues from human hepatocellular carcinoma (HCC) model on nude mice (established from high metastasis HCC cell line MHCC97LM6) were obtained. RESULTS: HfgI2 was detected in tumor tissues from 127 out of 133 patients as well as tumor tissues collected from human HCC nude mice. Hfgl2 was highly expressed both in cancer cells and interstitial inflammatory cells including macrophages, NK cells, and CD8^+ T lymphocytes and vascular endothelial cells. HfgI2 mRNA was localized in cells that expressed hfgI2 protein. Fibrin (nogen) colocalization with hfgl2 expression was determined by dual immunohistochemical staining. In vitro, IL-2 and IFN-γ, increased hfgl2 mRNA by 10-100 folds and protein expression in both THP-1 and HUVEC cell lines. One-stage clotting assays demonstrated that THP-1 and HUVEC cells expressing hfgl2 had increased procoagulant activity following cytokines stimulation. CONCLUSION: The hfgI2 contributes to the hypercoagulability in cancer and may induce tumor angiogenesis and metastasis via cytokine induction.
文摘目的:探讨纤维介素蛋白2凝血酶原酶(fibrinogen-like protein 2/fibroleukin,fgl2)在人肝癌(hepatocel-lular carcinoma,HCC)细胞系HCCLM6细胞的表达及在肿瘤生长中的作用。方法:用RT-PCR方法检测HCCLM6细胞在静息和细胞因子作用下fgl2mRNA水平的表达;用RNA干扰技术建立fgl2敲除的HCCLM6细胞模型;以流式细胞技术检测fgl2敲除后的HCCLM6细胞体外培养细胞周期和增殖指数的改变以及对TNF-α(肿瘤坏死因子-α)诱导凋亡的抵抗能力改变。结果:IFN-γ(干扰素-γ)、IL-1β(白介素-1β)和TNF-α均可在mRNA水平上调HCCLM6细胞fgl2的表达,以TNF-α诱导作用最为显著;fgl2干扰后HCCLM6细胞增殖指数显著降低,细胞周期分析S期比例显著下降;fgl2干扰对HCCLM6细胞在全培养基和无血清培养基中培养的凋亡率无显著影响,但在合并大剂量TNF-α诱导下,fgl2干扰可显著提高HCCLM6细胞的凋亡率。结论:炎性细胞因子可诱导HCCLM6细胞表达fgl2,fgl2的表达可直接促进肿瘤细胞的生长,并增强对TNF-α诱导凋亡的耐受。
文摘The aim of this study is to investigate the important regulative elements region which plays an important role on the activation of transcription exerted by the 5' noncoding region of hfgl2 gene in response to HBc and HBx. A series of promoter luciferase report plasmids, in which the hfgl2 gene has been deleted of the 5' and retained the common 3', were constructed. All the plasmids constructed were subjected to electrophoretic analysis and DNA sequencing. A eukaryotic construct expressing HBc or HBx, a luciferase reporter construct containing hfgl2 promoter and aβ-galactosidase (β-gal) plasmid were co-transfected into Chinese hamster ovary (CHO) cells and hepG2 cells, respectively. Luciferase report plasmids containing hfgl2 promoter were successfully constructed, and a serial assays of deletion of hfgl2 gene promoter showed that a strong regulatory region from -817 to -467 (relative to the transcription start site) was responsible for transcription and expression regulation of hfgl2 gene. The important regulative elements region in the promoter of hfgl2 gene was in response to HBc and HBx. which contributes to further pursuit of cis-acting elements and transcriptional factors involved in the transcription of hfgl2 gene.