Hepatoma-derived growth factor (HDGF) is a novel growth factor originally purified from media conditioned with the human hepatoma cell line HuH-7 Established studies have indicated: HDGF overexpression is related to poor prognosis in various types of cancer including melanoma. HDGF is composed of 240 amino acids and contains two bipartite nuclear location signals (NLSs). To date, the cell cycle distribution and function of HDGF remains largely uncharacterized. By arresting at various cell cycle stages, it was shown that the cellular HDGF level was highest in G1/S phase and lowest in M phase, which was inversely correlated with that of CDK1-CycB kinase. By using in vitro kinase assays, it was found that HDGF is phosphorylated by CDK1, but not glycogen synthase kinase 3b (GSK3b). Bioinformatic search predicted that Ser165 is the putative CDK1 phosphorylation site. Site-directed mutagenesis analysis found that Ser165Ala HDGF mutant was not phosphorylated by CDK1, suggesting Ser165 is indeed the phosphorylation site of CDK1. However, Ser165 mutation had no influence on nuclear targeting of HDGF. In protein stability assay using cyclohexamide, it was found that Ser165Ala mutant HDGF was more stable than the wild type (WT) HDGF. In addition, the HDGF phosphorylation by CDK1 in mitosis resulted in the ubiquitination and degradation of HDGF. Finally, Ser165Ala mutant HDGF was less potent than the WT HDGF in stimulating cell proliferation and migration. Together, these results indicate that CDK1 regulate the stability and function of HDGF.

Purpose: Hepatoma-derived growth factor (HDGF) is a novel growth factor that plays an important role in the pathogenesis and progression of a variety of cancers. The present study was designed to elucidate the role of HDGF expression in breast cancer. Patients and Methods: Tissues were collected from patients with breast cancer who underwent surgery. The expression of HDGF, Ki-67, FOXP3, p53, ER, PR and CD4+CD25+ in 71 patients with breast cancer containing a) malignant tissue (n = 58), b) uninvolved breast tissue obtained from tissue distant from the tumor (n = 13) using immunohistochemistry (IHC). The content of CD4+CD25 high in PBMC was determined by flow cytometry. Data were expressed as ROC Curve and the significance of the differences was assessed by ANOVA. Results: IHC analysis found that the expression level of HDGF and CD4+CD25high in tumor tissues was significantly higher than that in non-tumor tissues (P < 0.001). Besides, HDGF and CD4+CD25high expression was significantly correlated with tumor grades of breast cancer (P < 0.05). Increased circulating HDGF and CD4+CD25high levels were found in serum from patients with breast cancer compared with serum from healthy controls (P < 0.001). The nuclear HDGF labeling index was positively correlated with Ki-67, FOXP3 and p53 in breast cancer (P < 0.05). Finally, incubation with recombinant HDGF significantly increased the content of CD4(+)CD25high T cells in peripheral blood mononuclear cells (PBMCs). Conclusion: The present study demonstrated HDGF overexpression is correlated with tumor grades, recurrence, proliferation, and tumor immunity in breast cancer. In the future, HDGF may constitute a novel molecular target for diagnosis and treatment of breast cancer.

Hepatoma-derived growth factor (HDGF) is a novel growth factor identified from conditioned medium of hepatoma cell line. HDGF expression is upregulated in many types of cancer. Besides, HDGF it is well known that stimulate the proliferation and migration of various types including fibroblasts, hepatoma cells, and endothelial cells. HDGF is composed of 240 amino acids with a well conserved, N-terminal PWWP (conserved Pro-Trp-Trp-Pro motif) domain (residues 1-100) and a highly variable C-terminal domain (residues 101-240). PWWP domain binds to heparin and heparin sulfate located outside the surface of cell membrane and facilitated internalization of the protein into cells. There are two putative bipartite nuclear localization signals (NLSs) in HDGF. The cell surface receptor of HDGF is not been identified so far. The amino acids 81 to 100 are responsible for NIH3T3 membrane receptor binding domain and Lys96 played a major role in this domain. In the present study, I expressed and purified a functional recombinant HDGF protein from E. coli. Recombinant HDGF protein stimulated the growth and migration of cervical cancer HeLa cells. Subsequently, site-directed mutagenesis was been completed on produce recombinant HDGF protein with mutations in both NLSs and receptor binding (K96A). Respectively, Deletion of NLS1 and NLS2 abolished the nucleus targeting of HDGF and abrogated the growth promoting as well as the chemotaxic capability of recombinant HDGF. Substitution of a single amino acid (K96A) within this peptide was sufficient to diminish it is binding to the cell surface and it is proliferated activity. In summary, both NLSs and K96 residue are important to biological functions of HDGF protein.

Abstract PURPOSE: Hepatoma-derived growth factor (HDGF) is a unique nuclear/growth factor and plays an important role in the development and progression of cancer. The current study aimed to elucidate the correlation between HDGF expression, clinic-pathologic parameters, and associated molecular factors of oral cancer. MATERIALS AND METHODS: The surgically resected samples from a total of 95 patients with oral cancer (squamous cell carcinoma) were enrolled to construct the tissues microarray (TMA) in this retrospective study. The HDGF expression in TMA of oral cancer was determined by immunohistochemistry. HDGF and vascular endothelial growth factor (VEGF) immunostaining in tumor samples was scored and the labeling index were correlated with various clinic-pathologic parameters by statistic analysis. RESULTS: Expression of nuclear HDGF and VEGF was highly correlated with primary T stage (P=0.004 and P=0.038, respectively) and histological grade (P=0.013 and P=0.017, respectively). VEGF expression also associated with nodal status (P=0.021). Moreover, expression of nuclear HDGF and VEGF were highly correlated to each other (P=0.006). On the other hand, expression of HDGF in cytoplasm only associated with tumor necrosis (P=0.002) and showed no impact on survival. In univariate analysis, high expression of nuclear HDGF and VEGF significantly affected disease-specific, metastasis-free, and local recurrence-free survival. Multivariate analysis also indicated that expression level of nuclear HDGF is an independent prognostic factor for disease-specific and local recurrence-free survival (P=0.028; P=0.0285). Indeed, high expression of VEGF is also an independent factor in disease-specific, local recurrence-free, and metastasis free survival (P=0.0183; P=0.0461; P=0.0153). CONCLUSION: The data showed that high expression of nuclear HDGF and VEGF in squamous cell carcinoma of oral cavity might identify patients at risk of aggressive disease and predict poor prognosis. HDGF might play as key of regulation of tumorigenesis. Therefore, HDGF could be a candidate gene for the development of diagnostic and therapeutic strategies for oral cancer. Further studies are still need to determine the precise role of HDGF in the biological behavior of oral caner and the regulatory mechanism with other associated molecular factors.

Hepatoma-derived growth factor (HDGF) is a nucleus-targeting mitogen for various types of cells. Besides, HDGF overexpression is associated with tumor progression and poor survival outcome in patients with hepatocellular carcinoma (HCC) and lung cancer. HDGF is capable of promoting the proliferation and migration in various types of cells. HDGF is composed of 240 amino acids and contains 2 putative bipartite nuclear localization signals (NLSs). By dividing HDGF into two deletion domains: PWWP (residues 1-100) and C140 (residues 101-240), we found that both PWWP and C140 domains are capable of promoting nuclear localization, However, only C140 domain promoted cell proliferation and migration as HDGF. Mutation in NLS domains abrogated the nuclear localization and growth-promoting function, but not the migratory potential of HDGF. Beside, Ser165 was predicted as putative cdc2 phosphorylation site. In vitro kinase assay indicated that Ser165 of HDGF is the phosphorylated site of cdc2 kinase. We also demonstrated that mutations in cdc2 phoshprylation site did not affect the nuclear localization, proliferation-stimulating activities of HDGF but enhance migration-stimulating abilities of HDGF. Recently, the HDGF domain containing residues 81 to 100 is shown to be responsible for binding to membrane receptor in NIH3T3 cells. Besides, Lys96 plays a pivotal role for receptor binding. By generation of HDGF Lys96A mutant protein, we found that mutation of Lys96 indeed caused a prominent reduction in cellular binding affinity of HDGF to NIH3T3 cells and affect cell migration. In summary, the NLSs are essential for the mitogenic effect of HDGF, but not required for migration. And the cdc2 phoshorylation site is important for NIH3T3 migration. The Lys96 of HDGF play an important role of membrane receptor binding and cell migration.

Hepatocellular carcinoma (HCC) is the most common malignant tumor in Taiwan with more than one million new cases annually in the world. Growth factors play important roles in liver carcinogenesis. Hepatoma-derived growth factor (HDGF), originally isolated from the cultured media of human hepatoma HuH-7 cells, stimulate the growth of fibroblast cells, endothelial cells, and hepatoma cells. Overexpression of HDGF is related to the transformation of human hepatoma, lung cancer and melanoma. Besides, HDGF also exerts strong influence on the prognosis of patients with hepatoma. In this study, recombinant HDGF was expressed and purified with higher purity than 90%. The recombinant protein was used to raise polyclonal HDGF antibodies in rabbits and to generate three lines of HDGF monoclonal antibodies in mice. After antibodies characterization, an in-house, sandwich HDGF ELISA system was established using the purified polyclonal anti-HDGF IgG as the capture antibodies and the monoclonal anti-HDGF IgG as the detection antibodies. By using recombinant HDGF as standard, this ELISA system accurately evaluate the changes of HDGF release in SK-Hep-1 cells after gene delivery. In addition, we also evaluated the effect of anti-HDGF on the growth of hepatoma cells. Application of either polyclonal or monoclonal HDGF antibodies, but not preimmune antibodies, inhibited the proliferation of SK-Hep-1 hepatoma cells in a dose-dependent manner. In summary, the present study generated HDGF monoclonal antibodies for development of HDGF ELISA and application on suppressing HCC progression. Future studies should be carried out to enhance the sensitivity of HDGF ELISA and to evaluate the therapeutic potential of HDGF antibodies for treatment of HCC.

Hepatocellular carcinoma (HCC) is the most common and devastating malignant tumor in Taiwan. The major factors involved in the molecular pathogenesis for the development of HCC have been explored in recent years. An extensive array of growth factors and their receptors have been identified and may act as positive and negative modulators in different stages of hepatocarcinogenesis. HDGF (hepatoma-derived growth factor) is a novel growth factor, identified from conditioned medium of hepatoma cell line. HDGF has growth stimulating activity for fibroblast and some hepatoma cells. The high homology of protein sequence to HMG (high mobility group) protein but with distinct structure indicate it is a novel growth factor with mitogenic effect. Recently, elevated HDGF expression was found in developing kidneys but little in adult kidney. Besides, HDGF expression was found to be correlated with angiogenic status of tissues. Thus, it is speculated that HDGF plays a role during embryonic development and angiogenesis. Besides, HDGF also plays a role in cell-to-cell interaction and cell movement. HDGF is a growth factor that is involved in stimulating vascular smooth muscle cells (SMCs) proliferation during development and in disease. HDGF contains a true bipartite nuclear localization sequence necessary for nuclear targeting. It is required for HDGF stimulation of DNA replication and cell proliferation in vascular smooth muscle cell. In present studies, have transfected HDGF cDNA to 3T3 cell and the HDGF expression was verified by Western blot analysis. HDGF expression altered the morphologies and growth rate of 3T3 cells by 2-fold. Besides, HDGF-expressing cells were more resistant to serum-starvation. Injection of 3T3-HDGF cells, but not 3T3 cells, resulted in tumor formation in nude mice, suggesting that the angiogenic and mitogenic functions of HDGF might contribute to carcinogenesis. By using various reagents including H2O2, dexamethasome, taxol, cisplatin, CoCl2 and KCN, the cellular stress studies revealed differential responses between in 3T3 and 3T3-HDGFH. analyzed dose and time-dependent effects of UV irradiation and found that 3T3-HDGF cells are more sensitive to UV irradiation than 3T3 cells and susceptible to apoptosis. hope these experiments will bring further insights into the cellular function of HDGF, particularly during angiogenic process, thereby enable to evaluate its role during HCC progression and its potential as clinical marker and therapeutic target for HCC.

HDGF (hepatoma-derived growth factor) is a novel growth factor,identified from conditioned medium of hepatoma cell line. HDGF has growth stimulating activity for fibroblast and some hepatoma cells. HDGF, a novel defined growth factor with mitogenic effect, has homology protein sequence as HMG (high mobility group) protein and their three dimension structures appeared to be similar to each other. Recently, elevated HDGF expression was found in developing kidneys but less was found in adult kidney. In addition, HDGF expression was found to be correlated with angiogenic status of tissues. Thus, it is speculated that HDGF plays a role during embryonic development and angiogenesis. HDGF also plays a role in cell-cell interaction and cell migration. HDGF is a growth factor that is involved in stimulating vascular smooth muscle cells (SMCs)proliferation during development and in disease. HDGF contains a true bipartite nuclear localization sequence necessary for nuclear targeting. HDGF is sciential factor in stimulating DNA replication and cell proliferation of vascular smooth muscle cell.In this study,we used E. coli strain BL21 (DE3) to express the recombinant protein hepatoma derived growth factor(HDGF). To find out the optimal production conditions,we studied on the different temperature and fermentor to calculate all cost .

Le neuroblastome est un cancer pédiatrique diagnostiqué chez les très jeunes enfants. afin de pouvir étudier l'impact de signalisation embryonnaire sur le développement de la maladie in vivo, nous avons mis au point un nouveau modèle d'étude du neuroblastome chez l'embryon aviaire. Ce nouveau modèle nous permet d'étudier in vivo la contribution de différentes signalisation à la tumorigénèse neuroblastique. nous avons donc mis en évidece le rôle de Sema3C et HDGF dans la tumorigénèse neuroblastique / Neuroblastoma is a pediatric cancer diagnosed in very young children. In order to study the impact of embryonic signaling on the development of the disease in vivo, we have developed a new model for the study of neuroblastoma in the avian embryo. This new model allows us to study in vivo the contribution of different signaling to neuroblastic tumorigenesis. We have thus highlighted the role of Sema3C and HDGF in neuroblastic tumorigenesis

Cutaneous malignant melanoma is the fastest increasing malignancy in humans. Hepatoma-derived growth factor (HDGF) is a novel growth factor identified from human hepatoma cell line. HDGF overexpression is correlated with poor prognosis in various types of cancer including melanoma. However, the underlying mechanism of HDGF overexpression during melanoma carcinogenesis remains unclear. In this study, adding exogenous HDGF stimulated the invasion and colonies formation of B16-F10 melanoma cells. Adenovirus vectors encoding HDGF and HDGF-RNAi were generated and characterized to up- and down-regulated HDGF expression in B16-F10 melanoma cells. It was found that HDGF overexpression stimulated the proliferation, invasiveness, anchorage-independent growth of B16-F10 melanoma cells whereas HDGF knockdown exerted opposite effects. In lung-metastasis model, intravenous injection of HDGF-overexpressing melanoma cells resulted in increased metastasis while HDGF-downregulated melanoma cells caused decreased metastasis. Similarly, in primary melanoma model, subcutaneous injection of HDGF-overexpressing melanoma cells enhanced while HDGF-downregulated melanoma cells reduced the tumor burden in mice. Histological analysis revealed increased tumor proliferation and neovascularization with concomitant reduction of apoptosis in HDGF-overexpressing melanoma. Moreover, HDGF-overexpressing melanoma also exhibited enhanced propensity to metastasize from the primary tumors to lymph node and lung. Finally, it was found that HDGF overexpression increased nuclear factor kappa B (NF£eB) activities and Akt phosphorylation up and down stream alternation like PI3K, PTEN, I£eB and it¡¦s subunit IKK£\, IKK£], IKK£^ in melanoma cells. It also found that HDGF overexpression influenced MITF and HIF1£\ in melanoma after gene delivery. HDGF also altered EMT changes like E,N-cadherin, vimentin, and £],£^-catenin. The present study provides conclusive evidence that HDGF upregulation promotes the growth and metastasis of melanoma by promoting the survival and vascularization. Besides, HDGF knockdown may constitute a novel strategy for melanoma control.