Hepatocyte Growth Factor Concentrations Research Articles

Hepatocyte growth factor — the earliest marker of myocardial injury in ST-segment elevation myocardial infarction

Hepatocyte growth factor (HGF) concentration increases in the first few hours of myocardial infarction (MI). (1) To illustrate human HGF (hHGF) plasma concentration during the first 24 h of ST segment elevation myocardialinfarction (STEMI); (2) To estimate the odds ratio of STEMI in the context of hHGF measurements; (3) To describe the normalconcentration of hHGF in healthy subjects. The study groups consisted of 73 STEMI patients and 11 healthy volunteers. In all the patients, we took bloodsamples for hHGF twice, i.e. on admission to hospital and 24 h later. The median value of hHGF in healthy volunteers was 666 pg/mL (576; 760 pg/mL). In STEMI, the highest values of hHGF were observed in the first measurement. An increase of 1 pg/mL in hHGF level increased STEMI odds ratio by 0.2%. In acute MI, of the known biomarkers, hHGF rises the earliest and very promptly returns to normal values.

Exercise and the platelet activator calcium chloride both influence the growth factor content of platelet-rich plasma (PRP): overlooked biochemical factors that could influence PRP treatment

BackgroundThere is strong evidence that exercise affects platelet haemostasis factors, but this potential effect on growth factor concentrations in platelet-rich plasma (PRP) has never been studied. In addition, there is...

The effect of cabergoline on folicular microenviroment profile in patients with high risk of OHSS

The aim of this study to evaluate the effect of cabergoline on follicular microenvironment by measuring follicular fluid (FF) insulin like growth hormone –I (IGF-I), antimullerian hormone (AMH), inhibin B and hepatocyte growth factor (HGF) levels in women with PCOS and high risk of ovarian hyperstimulation syndrome (OHSS). In this prospective cohort study, 41 women with PCOS undergoing controlled ovarian hyperstimulation for assisted reproduction and having the high risk factors for OHSS are included. The women in the study group (n = 15) received cabergoline for OHSS prevention while the women in the control did not received any medications for OHSS prevention. FF samples were collected during oocyte pick-up procedure for all women were determined using commercially available ELISA kits. Concentrations of FF IGF-I, AMH, inhibin B and HGF were assessed. In the study group FF AMH (2.96 ± 1.27 versus 1.91 ± 0.64 ng/mL), Inhibin B (1339.47 ± 198.56 versus 1200.09 ± 133.64 pg/mL), HGF (5623.21 ± 2411.09 versus 3787.42 ± 2269.89 pg/mL) and IGF-I (298.60 ± 37.80 versus 219.90 ± 71.40 pg/mL) concentrations were significantly decreased compared with control group. Cabergolin prevents OHSS in high risk patients by disrupting FF hormone microenvironment.

Cancer-Associated Fibroblasts from Hepatocellular Carcinoma Promote Malignant Cell Proliferation by HGF Secretion

Cancer-associated fibroblasts (CAFs) are reported to support tumorigenesis by stimulating angiogenesis, cancer cell proliferation, and invasion in most solid tumors. However, the roles of CAFs in the liver cancer microenvironment have not been thoroughly studied. In our previous study, we successfully isolated CAFs from hepatocellular carcinoma (HCC) (H-CAFs) and proved that H-CAFs suppressed the activation of NK cells and thereby created favorable conditions for HCC progression. In our present study, we found that the proliferation of MHCC97L and Hep3B cells was significantly promoted by treatment with conditioned medium from H-CAFs. Pathological analysis also revealed that H-CAFs increased the proportion of Ki-67 (+) malignant cells and prevented them from undergoing necrosis. Moreover, the concentration of hepatocyte growth factor (HGF) cytokine in the conditioned medium of H-CAFs was higher than conditioned medium from normal skin fibroblasts (NSFs). Anti-HGF significantly reduced the proliferation-promoting capability of H-CAFs. In addition, we found that the abundance of H-CAFs correlated positively with tumor size. These results indicate that H-CAFs are an important factor for promoting the growth of HCC in vitro and in vivo, and that HGF plays a key role in HCC proliferation induced by H-CAFs.

Abstract 1627: Fibroblast HGF elicits c-MET-mediated signaling and migration in ovarian cancer cells.

Abstract Background: Epithelial ovarian cancer (EOC) has the highest mortality rate of all gynecologic malignancies diagnosed in the U.S. due to its rapid progression without symptom. Stromal components in the ovary contribute to rapid progression of cancer and interruption of their interaction with cancer cells are being explored as important therapeutic target. Hepatocyte growth factor (HGF) is expressed primarily in mesenchymal cells and acts on epithelial cells. HGF is also a unique cytokine that induces pleiotropic biological responses, involving proliferation, invasion, and dissemination of cancer cells. We studied c-MET/HGF axis as a mediator of tumor-stromal interactions in ovarian cancer using human ovarian fibroblast and their derived extracellular matrices (ECM). Methods: Six EOC cells were each cultured alone or in the presence of three human ovarian fibroblast cultures and their derived ECM in Transwell plates and the number of migrated cells was quantitated. ELISA was used to measure the concentration of HGF secreted by cancer cells and fibroblasts or released from fibroblast-derived ECM. EOC cells were treated with conditioned media derived from fibroblast that secretes high level of HGF and c-MET mediated signalling was assessed by immunoblotting. Results: Five out of the six EOC cells tested highly expressed c-MET and none produced detectable levels of HGF. IHFNO-303, a human ovarian fibroblast line, secreted the highest level of HGF (∼3,800 pg/mL) and markedly enhanced migration (2 to 140-fold increase) of the EOC cells that express c-MET. Recombinant HGF and a HGF neutralizing antibody were used to validate HGF as a major migratory factor secreted by fibroblast. HGF sequestered within the fibroblast-derived ECM was also able to stimulate cell migration by 1.5 to 24-fold, when released via EOC-associated degradation of the ECM. In cells containing constitutive c-MET phosphorylation, recombinant HGF and fibroblast-derived HGF negligibly affect c-MET phosphorylation on Tyr1234 and Tyr1003. However, both sources of HGF increased the phosphorylation of c-MET on Tyr1349, the multisubstrate docking site, by more than 6-fold and led to activation of downstream signalling transducers, e.g., AKT and ERK, in all c-MET expressing EOC cells. Therefore, signalling mediated by c-MET and HGF likely occurs in a paracrine manner in ovarian cancer. Conclusion: Our study demonstrates the functional contribution of c-MET and HGF signalling in malignant progression driven by important tumor-stromal interactions. We demonstrate for the first time that phosphorylation of c-MET on Tyr1349 requires extraneous HGF and phosphorylated c-MET level on Try1349 might be a good indicator of c-MET activation in the microenvironment and likely responsive to c-MET targeted therapy. Citation Format: Youngjoo Kwon, Yan Zhou, Andrew K. Godwin. Fibroblast HGF elicits c-MET-mediated signaling and migration in ovarian cancer cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1627. doi:10.1158/1538-7445.AM2013-1627

Abstract 2374: Tumoral MET expression and plasma HGF level in patients with ALK 2p23 fusion driven lung cancer.

Abstract Background. MET is the tyrosine kinase receptor for the ligand hepatocyte growth factor (HGF). MET kinase has been identified as a novel promising target in non-small cell lung lung cancer (NSCLC). Crizotinib, a recently FDA-approved ALK inhibitor for NSCLC harboring ALK 2p23 fusion (ALK+), was initially developed as a MET inhibitor. MET expression level is a potential biomarker for response to MET inhibitors. The role of MET/HGF pathway in ALK+ NSCLC is unknown. Methods. A cohort of 51 NSCLC patients tested for ALK fusion at Cleveland Clinic (2000 to 2012), including 15 ALK(+) and 36 ALK(-) subjects, were included in the study. Immunohistochemistry (IHC) stain against MET was performed using a monoclonal CONFIRM anti-Total c-MET antibody (SP44, Ventana). Staining was performed on a Ventana Benchmark XT automated immunostainer (Ventana Medical Systems). A four-tier (0, 1+, 2+, 3+) scoring system and H-score (the sum of each intensity score x %) were used. Thirteen ALK(+) patients treated on crizotinib therapy were prospectively followed and a total of 32 blood plasma samples were collected (1) prior to crizotinib initiation, (2) during responding period and (3) at disease progression, for HGF level measurement using HGF ELISA kit (R&D). Statistical analysis was performed using Fisher exact test and Wilcoxon rank sum test in JMP. Results. Of the 51 tumor tested for MET IHC, 39 were adenocarcinoma (15 ALK+ and 24 ALK-), 6 squamous cell, 4 large cell carcinoma and 2 other NSCLC. None received any MET inhibitor when tissue was collected. The percentage of negative MET IHC (score 0 or 1+ in 50% tumor cells) was not statistically significant between ALK(+) and ALK(-) groups (40% vs 30%, p=0.5). The plasma HGF levels in ALK(+) patients varies from undetectable to 14000pg/ml. The median (25th-75th quantiles) level in patients prior to crizotinib therapy (n=5), during responding period (CR/PR/SD, n=12) and at disease progression (n=5) were 2137(767-12100)pg/ml, 1629(785-3430)pg/ml and 4064(1108-7308)pg/ml. The plasma HGF level showed a trend of decreasing during clinical response to crizotinib and elevation upon disease progression, albeit not reaching statistical significance in this small study cohort (p>0.05). Conclusions. MET expression is commonly seen in NSCLC, and no significant difference were observed between fusion ALK(+) and ALK(-) NSCLC. The baseline plasma HGF concentration varied significantly among ALK(+) NSCLC patients, but showed a trend correlating with therapeutic response and resistant progression, to crizotinib therapy. Prospective study with larger cohorts is warranted to further evaluate the role of MET/HGF as biomarker in ALK targeted therapy. Citation Format: Yan Feng, Eugen Minca, Wei Zhang, Lihong Yin, Nathan Pennell, Carol Farver, Raymond Tubbs, Angen Liu, Patrick Ma. Tumoral MET expression and plasma HGF level in patients with ALK 2p23 fusion driven lung cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2374. doi:10.1158/1538-7445.AM2013-2374

Conditional genetic elimination of hepatocyte growth factor in mice compromises liver regeneration after partial hepatectomy.

Hepatocyte growth factor (HGF) has been shown to be indispensable for liver regeneration because it serves as a main mitogenic stimulus driving hepatocytes toward proliferation. We hypothesized that ablating HGF in adult mice would have a negative effect on the ability of hepatocytes to regenerate. Deletion of the HGF gene was achieved by inducing systemic recombination in mice lacking exon 5 of HGF and carrying the Mx1-cre or Cre-ERT transgene. Analysis of liver genomic DNA from animals 10 days after treatment showed that a majority (70–80%) of alleles underwent cre-induced genetic recombination. Intriguingly, however, analysis by RT-PCR showed the continued presence of both unrecombined and recombined forms of HGF mRNA after treatment. Separation of liver cell populations into hepatocytes and non-parenchymal cells showed equal recombination of genomic HGF in both cell types. The presence of the unrecombined form of HGF mRNA persisted in the liver in significant amounts even after partial hepatectomy (PH), which correlated with insignificant changes in HGF protein and hepatocyte proliferation. The amount of HGF produced by stellate cells in culture was indirectly proportional to the concentration of HGF, suggesting that a decrease in HGF may induce de novo synthesis of HGF from cells with residual unrecombined alleles. Carbon tetrachloride (CCl4)-induced regeneration resulted in a substantial decrease in preexisting HGF mRNA and protein, and subsequent PH led to a delayed regenerative response. Thus, HGF mRNA persists in the liver even after genetic recombination affecting most cells; however, PH subsequent to CCl4 treatment is associated with a decrease in both HGF mRNA and protein and results in compromised liver regeneration, validating an important role of this mitogen in hepatic growth.

Efficient generation of functional hepatocyte-like cells from menstrual blood-derived stem cells

In recent years, the advantages of menstrual blood-derived stem cells (MenSCs), such as minimal ethical considerations, easy access and high proliferative ability, have inspired scientists to investigate the potential of MenSCs in cell therapy of different diseases. In order to characterize the potency of these cells for future cell therapy of liver diseases, we examined the potential of MenSCs to differentiate into hepatocytes, using different protocols. First, the immunophenotyping properties and potential of MenSCs to differentiate into osteoblasts, adipocytes and chondrocytes were evaluated. Thereafter, the differentiation protocols developed by two concentrations of hepatocyte growth factor (HGF) and oncostatin M (OSM), in combination with other components in serum-supplemented or serum-free culture media, were also investigated. The sequential differentiation was monitored by real-time PCR, immunostaining and functional assays. Our primary data revealed that the isolated MenSCs exhibited mesenchymal stem cell markers in parallel to OCT-4 as an embryonic marker. Regardless of differentiation procedures, the developed cells expressed mature hepatocyte markers, such as albumin, tyrosine aminotransferase and cytokeratin-18 at the mRNA and protein levels. They also showed functional properties of hepatocytes, including albumin secretion, glycogen storage and cytochrome P450 7A1 expression. However, the degree of differentiation was dependent on the concentrations of HGF and OSM. Indeed, omission of serum during the differentiation process caused typical improvement in hepatocyte-specific functions. This study is a novel report demonstrating the differentiation potential of MenSCs into hepatocyte-like cells. We recommend a complementary serum-free differentiation protocol for enrichment of in vitro production of functional MenSC-derived hepatocyte-like cells that could lead to a major step toward applied stem cell therapy of chronic liver diseases.

Hepatocyte growth factor and interferon‐γ inducible protein‐10 are related to visceral adiposity

Increased production of chemokines by adipose tissue and defective adipose tissue oxygenation as a result of obesity may induce leucocyte infiltration and subsequent systemic inflammation. 1-To determine the relation between the amount of visceral and subcutaneous adipose tissue and the chemokine interferon-γ-inducible protein 10 (IP-10) and angiogenic factor hepatocyte growth factor (HGF). 2-To determine the relation between the metabolic syndrome and IP-10 as well as HGF. Patients originated from the Secondary Manifestations of ARTerial disease (SMART) cohort. In this study, a cohort of 1251 patients with manifest vascular disease was included. Subcutaneous and visceral adipose tissue thickness (SAT and VAT respectively) were measured ultrasonographically. IP-10 and HGF concentrations were measured with Luminex multiplex immuno assay in addition to fasting metabolic parameters. Linear regression analyses with adjustments for age, gender, smoking, estimated glomerular filtration rate, type 2 diabetes mellitus and medication use were applied to quantify the relations between adiposity or metabolic syndrome and IP-10 and HGF concentrations. VAT was significantly associated with (log)IP-10 and (log)HGF, reflected by significant higher β-values in VAT quartile 4 compared with VAT quartile 1 (reference): β0.155 (95%CI:0.073-0.237) for IP-10 and β0.147 (95%CI:0.076-0.218) for HGF. Per standard deviation increase in VAT, (log)IP-10 levels increased with 0.057 pg/mL (95%CI:0.027-0.087) and (log)HGF increased with 0.051 pg/mL (95%CI:0.025-0.077). Effect estimates were not affected by including body mass index(BMI) in the model. In contrast, SAT was not associated with IP-10 and HGF. Furthermore, the presence of the metabolic syndrome was associated with IP-10 and HGF. Visceral adipose tissue but not subcutaneous adipose tissue is significantly associated with circulating levels of IP-10 and HGF, irrespective of BMI.

Differences in the expression of hepatocyte growth factor in acute and chronic bowel inflammation—Implications for diagnosis?

Background: Hepatocyte growth factor (HGF) acts as an acute phase protein with regenerative properties. HGF is produced systemically and locally during inflammation but exhibits decreased binding affinity to heparan sulphate proteoglycan (HSPG)/glycosaminoglycan during chronic inflammation. We previously observed a high faecal concentration and binding affinity of HGF to HSPG during acute gastroenteritis. High faecal concentrations of calprotectin and HGF have been reported in chronic inflammatory bowel disease (IBD). Methods: Stool samples from patients with ulcerative colitis in remission (n = 11) or exacerbation (n = 5), microscopic colitis (n = 11), colon cancer (n = 6), or acute gastroenteritis caused by Clostridium difficile (n = 20), as well as healthy controls (n = 7), were analysed for the presence of HGF by ELISA, surface plasmon resonance, SDS-PAGE, and Western blot. Then in two patients with ulcerative colitis exacerbation and C. difficile infection, the expression of HGF and calprotectin was studied in colonic biopsies. Results: The faecal concentration of HGF was significantly higher in patients with ulcerative colitis compared to the other groups. The binding affinity to dextran was lower in all groups compared to acute inflammation. HGF receptor binding was similar across groups. In a patient with concomitant C. difficile infection and distal ulcerative colitis, HGF was highly expressed in the part of the bowel unaffected by ulcerative colitis, but no expression was found at the site of chronic inflammation. In the patient with total colitis the biopsies showed low expression of HGF. The areas with chronic inflammation exhibited infiltrating calprotectin-stained neutrophils. Conclusion: HGF is produced locally during inflammation of the bowel. The HGF produced during acute inflammation or exacerbations of chronic inflammation by the unaffected area shows binding affinity to glucosaminoglycans. Measuring HGF binding in faeces and biopsies may be a tool for differentiating between acute and chronic bowel inflammation, which should be assessed thoroughly in future studies.

Human platelets inhibit liver fibrosis in severe combined immunodeficiency mice

To investigate the role of human platelets in liver fibrosis. Severe combined immunodeficiency (SCID) mice were administered CCl4 and either phosphate-buffered saline (PBS group) or human platelet transfusions (hPLT group). Concentrations of hepatocyte growth factor (HGF), matrix metallopeptidases (MMP)-9, and transforming growth factor-β (TGF-β) in the liver tissue were compared between the PBS and the hPLT groups by enzyme-linked immunosorbent assay (ELISA) and Western blotting. The effects of a human platelet transfusion on liver fibrosis included the fibrotic area, hydroxyproline content, and α-smooth muscle actin (α-SMA) expression, which were evaluated by picrosirius red staining, ELISA, and immunohistochemical staining using an anti-mouse α-SMA antibody, respectively. Phosphorylations of mesenchymal-epithelial transition factor (Met) and SMAD3, downstream signals of HGF and TGF-β, were compared between the two groups by Western blotting and were quantified using densitometry. Hepatocyte apoptosis was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling. Furthermore, the accumulation of human platelets in the liver 2 h after platelet transfusion was compared between normal and fibrotic livers by immunohistochemical staining using an anti-human CD41 antibody. The fibrotic area and hydroxyproline content in the liver were both significantly lower in the hPLT group when compared to the PBS group (fibrotic area, 1.7% ± 0.6% vs 2.5% ± 0.6%, P = 0.03; hydroxyproline content, 121 ± 26 ng/g liver vs 156 ± 47 ng/g liver, P = 0.04). There was less α-smooth muscle actin staining in the hPLT group than in the PBS group (0.5% ± 0.1% vs 0.8% ± 0.3%, P = 0.02). Hepatic expression levels of mouse HGF and MMP-9 were significantly higher in the hPLT group than in the PBS group (HGF, 109 ± 13 ng/g liver vs 88 ± 22 ng/g liver, P = 0.03; MMP-9, 113% ± 7%/GAPDH vs 92% ± 11%/GAPDH, P = 0.04). In contrast, the concentration of mouse TGF-β in the liver tissue was significantly lower in the hPLT group than in the PBS group (22 ± 5 ng/g liver vs 39 ± 6 ng/g liver, P = 0.02). Phosphorylation of Met was more prevalent in the hPLT group than in the PBS group (37% ± 4%/GAPDH vs 20% ± 8%/GAPDH, P = 0.03). Phosphorylation of SMAD3 was weaker in the hPLT group than in the PBS group (60% ± 12%/GAPDH vs 84% ± 12%/GAPDH, P = 0.1), although this difference was not significant. Furthermore, a lower rate of hepatocyte apoptosis was observed in the hPLT group than in the PBS group (5.9% ± 1.7% vs 2.9% ± 2.1%, P = 0.02). Significant human platelet accumulation was observed in the fibrotic liver tissues, whereas few platelets accumulated in the normal liver. Human platelets inhibit liver fibrosis in SCID mice. Increased concentration of HGF in the liver suppresses hepatic stellate cell activation, induces MMPs, and inhibits hepatocyte apoptosis.

Decreased Hepatocyte Growth Factor (HGF) and Gamma Aminobutyric Acid (GABA) in Individuals with Obsessive-Compulsive Disorder (OCD)

IntroductionThere is support for the role of gamma aminobutyric acid (GABA) in the etiology of mood disorders. Recent research has shown that hepatocyte growth factor (HGF) modulates GABAergic inhibition and seizure susceptibility. This study was designed to determine and correlate plasma levels of HGF and GABA as well as symptom severity in individuals with obsessive-compulsive disorder (OCD).Subjects and methodsPlasma from 15 individuals with OCD (9 males, 6 females;, mean age 38.7 years) and 17 neurotypical controls (10 males, 7 females; mean age 35.2 years) was assessed for HGF, GABA, urokinase plasminogen activator (uPA), and urokinase plasminogen activator receptor (uPAR) concentration using enzyme-linked immunosorbest assays ELISAs. Symptom severity was assessed in these OCD individuals and compared with HGF and GABA concentrations.ResultsIn this preliminary study, individuals with OCD had significantly decreased HGF levels, decreased plasma levels of GABA and decreased uPA. We found that both uPA and uPAR levels correlate with HGF. Both low uPA and low uPAR levels correlate with high symptom severity in individuals with OCD. Low GABA levels in OCD individuals also correlate with high symptom severity.DiscussionThese results demonstrate a preliminary association between HGF, GABA, uPA levels, and OCD and suggest that plasma GABA and uPA levels are related to symptom severity in individuals with OCD.

Correlation between Hepatocyte Growth Factor (HGF) and Gamma-Aminobutyric Acid (GABA) Plasma Levels in Autistic Children

There is much support for the role of Gamma-Aminobutyric acid (GABA) in the etiology of autism. Recent research has shown that hepatocyte growth factor (HGF) modulates GABAergic inhibition and seizure susceptibility. This study was designed to determine and correlate plasma levels of HGF, GABA, as well as symptom severity, in autistic children and neurotypical controls. Plasma from 48 autistic children and 29 neurotypical controls was assessed for HGF and GABA concentration using ELISAs. Symptom severity was assessed in these autistic individuals and compared to HGF and GABA concentrations. We previously reported that autistic children had significantly decreased levels of HGF. In this study, the same autistic children had significantly increased plasma levels of GABA (P = 0.002) and decreased HGF levels correlated with these increased GABA levels (r = 0.3; P = 0.05). High GABA levels correlated with increasing hyperactivity (r = 0.6; P = 0.0007) and impulsivity severity (r = 0.5; P = 0.007), tip toeing severity (r = 0.35; P = 0.03), light sensitivity (r = 0.4; P = 0.02), and tactile sensitivity (r = 0.4; P = 0.01). HGF levels did not correlate significantly with any symptom severity. These results suggest an association between HGF and GABA levels and suggest that plasma GABA levels are related to symptom severity in autistic children.

Maternal Plasma Hepatocyte Growth Factor Concentrations in Women Who Subsequently Developed Preeclampsia

and remits dramatically after the placenta has been delivered. Despite extensive research, the pathophysiology of this condition remains unclear. Accumulating evidence, however, indicates that failure of the cytotrophoblast invasion of maternal uterine spiral arteries plays a central role. 4) Consequently, reduction of utero placental blood perfusion by shallow implantation results in local placental hypoxia. Chronic hypoxia Maternal Plasma Hepatocyte Growth Factor Concentrations in Women Who Subsequently Developed Preeclampsia

Abstract A20: Predictive value of plasma hepatocyte growth factor levels in gastric cancer risk: A Nested Case-Control Study within the Korean Multicenter Cancer Cohort

Abstract Background: HGF (hepatocyte growth factor) is also known as a scatter factor and is multifunctional cytokine that modulates processes such as growth, adhesion, motility, matrix degradation, invasion, and angiogenesis. We investigated whether serum HGF was a biomarker for the early detection and diagnosis of gastric cancer in a nested case-control study. Methods: The study population was composed of 282 cases and 282 matched controls within the Korean Multicenter Cancer Cohort. Plasma HGF was measured by enzyme-linked immunosorbent assay (ELISA). Conditional logistic regression models were used to compute the odds ratio (OR) and 95% confidence intervals (CI). Results: According to stratified analysis on age, sex, H. pylori infection, CagA and smoking, gastric cancer patients which are older, female, H. pylori positive, CagA positive and non-smokers had significantly higher serum HGF concentrations than normal controls. When conducting a logistic regression analysis of a group with the lowest concentration as a reference after dividing them into quartile according to serum HGF concentration, also, a group with the highest serum HGF concentration(≥393.7 pg/mL) showed a significantly increased risk of gastric cancer(OR=2.4 95% CI 1.4-3.9). In addition, higher serum HGF concentrations were associated with an increased risk for gastric cancer and it was statistically significant (p for trend=0.0054). Conclusions: Our finding suggested the possibility that HGF protein can be a biomarker to predict the risk of gastric cancer. Citation Format: Seung Hyun Ma, Jae Jeong Yang, Ko Kwang-Pil, Aesun Shin, Hai-Rim Shin, Daehee Kang, Keun-Young Yoo, Sue K. Park. Predictive value of plasma hepatocyte growth factor levels in gastric cancer risk: A nested case-control study within the Korean Multicenter Cancer Cohort. [abstract]. In: Proceedings of the Eleventh Annual AACR International Conference on Frontiers in Cancer Prevention Research; 2012 Oct 16-19; Anaheim, CA. Philadelphia (PA): AACR; Cancer Prev Res 2012;5(11 Suppl):Abstract nr A20. Note: This abstract was not presented at the conference.

Trametinib for patients with advanced melanoma – Authors' reply

Gerald Falchook and colleagues show clinical activity of the MEK inhibitor trametinib in patients with either BRAF-mutant or wild-type melanoma. Recently, two essential topics for melanoma have been reported: hepatocyte growth factor (HGF)-dependent resistance to therapy and new driver mutations in melanoma. Stroma-mediated resistance to treatment of BRAF-mutant melanoma is common and HGF secretion from stromal cells seems to be responsible for this drug resistance. Analysis of biopsy samples from patients with BRAF-mutant melanoma suggests that patients with abundant HGF from stromal cells have poor responses to treatment. Furthermore, an inverse relation has been reported between plasma HGF concentration and response to treatment in patients with BRAF-mutant melanoma. HGF secretion leads to activation of the HGF receptor MET, and dual inhibition of RAF and either HGF or MET results in reversal of drug resistance in BRAFmutant melanoma. In Falchook and colleagues’ study, examination by immuno histochemistry of whether HGF expression in melanoma sections correlated with poor responses to the MEK inhibitor trametinib would have been informative, not only in patients with the BRAF mutation, but also in patients with wild-type BRAF. These results would clarify the possibility that a combination of the MEK inhibitor and MET inhibitors have a synergistic eff ect. Moreover, although EGF is less able than HGF to reactivate ERK in most melanoma cell lines, EGF and EGFR mediated resistance might be of interest in melanoma and in colon cancers. For driver mutations, six genes related to melanoma have been newly identifi ed (PPP6C, RAC1, SNX31, TACC1, STK19, and ARID2), with RAC1 mutations also reported. The Illumina platform was used in Falchook and colleagues’ study to analyse the mutational and copy number status of 78 diff erent genes commonly implicated in tumorigenesis. Within these data, is there any information about the six new melanoma genes? If these driver mutations are present, examination of whether they correlate with reduced responses to therapy with trametinib would be interesting, because such examination would not only contribute to drug response predictions but also might help select other treatment options, including combination therapies. Furthermore, data on amplifi cation of BRAF, either with or without mutations, could provide information on whether copy number of this gene plays a role in response to therapy? In the context of treatment with the MEK inhibitor trametinib, an integrated examination including HGF secretion and other gene mutations would open up the possibility of prediction of drug response and combination therapy.

Trametinib for patients with advanced melanoma

Gerald Falchook and colleagues show clinical activity of the MEK inhibitor trametinib in patients with either BRAF-mutant or wild-type melanoma. Recently, two essential topics for melanoma have been reported: hepatocyte growth factor (HGF)-dependent resistance to therapy and new driver mutations in melanoma. Stroma-mediated resistance to treatment of BRAF-mutant melanoma is common and HGF secretion from stromal cells seems to be responsible for this drug resistance. Analysis of biopsy samples from patients with BRAF-mutant melanoma suggests that patients with abundant HGF from stromal cells have poor responses to treatment. Furthermore, an inverse relation has been reported between plasma HGF concentration and response to treatment in patients with BRAF-mutant melanoma. HGF secretion leads to activation of the HGF receptor MET, and dual inhibition of RAF and either HGF or MET results in reversal of drug resistance in BRAFmutant melanoma. In Falchook and colleagues’ study, examination by immuno histochemistry of whether HGF expression in melanoma sections correlated with poor responses to the MEK inhibitor trametinib would have been informative, not only in patients with the BRAF mutation, but also in patients with wild-type BRAF. These results would clarify the possibility that a combination of the MEK inhibitor and MET inhibitors have a synergistic eff ect. Moreover, although EGF is less able than HGF to reactivate ERK in most melanoma cell lines, EGF and EGFR mediated resistance might be of interest in melanoma and in colon cancers. For driver mutations, six genes related to melanoma have been newly identifi ed (PPP6C, RAC1, SNX31, TACC1, STK19, and ARID2), with RAC1 mutations also reported. The Illumina platform was used in Falchook and colleagues’ study to analyse the mutational and copy number status of 78 diff erent genes commonly implicated in tumorigenesis. Within these data, is there any information about the six new melanoma genes? If these driver mutations are present, examination of whether they correlate with reduced responses to therapy with trametinib would be interesting, because such examination would not only contribute to drug response predictions but also might help select other treatment options, including combination therapies. Furthermore, data on amplifi cation of BRAF, either with or without mutations, could provide information on whether copy number of this gene plays a role in response to therapy? In the context of treatment with the MEK inhibitor trametinib, an integrated examination including HGF secretion and other gene mutations would open up the possibility of prediction of drug response and combination therapy.

Hepatocyte Growth Factor and E-Cadherin in Acute Pancreatitis. Time Course and Early Assessment of Disease Severity

Objectives To evaluate the hepatocyte growth factor (HGF) and E-cadherin (EC) time course in the early phases of acute pancreatitis and to explore the usefulness of these molecules in assessing the severity of the disease. Patients Sixteen consecutive acute pancreatitis patients (7 males, 9 females; mean age 62.8 years, range 30-79 years) admitted to the hospital within 6 hours after the onset of pain; 16 age- and sex-matched healthy subjects were also studied. Eleven patients had mild pancreatitis and 5 had a severe disease. The acute pancreatitis was of biliary origin in 7 patients (43.8%), due to ERCP in 4 (25.0%), due to alcohol abuse in 2 (12.5%) and idiopathic in the remaining 3 (18.8%). Methods HGF and EC were quantified on hospital admission and for the following 2 days by using commercially available kits (Human HGF Immunoassay and Human sE-Cadherin Immunoassay; RD P=0.015) and the second day (57.7±25.2 ng/mL; P=0.015) of the disease than in healthy subjects (mean±SD: 40.5±11.6 ng/mL). HGF and EC serum concentrations were not significantly different between patients with severe disease and those with mild disease (P>0.089). Conclusions HGF and EC reflect an inflammatory response in acute pancreatitis patients; however, serum levels of these two molecules are not related to the severity of acute pancreatitis.

Enhanced PKCδ and ERK Signaling Mediate Cell Migration of Retinal Pigment Epithelial Cells Synergistically Induced by HGF and EGF

Proliferative vitreoretinopathy (PVR) and proliferative diabetic retinopathy (PDR) are characterized by the development of epi-retinal membranes which may exert a tractional force on retina. A lot of inflammatory growth factors may disturb the local ocular cells such as retinal pigment epithelial (RPE) cells, causing them to migrate and proliferate in the vitreous cavity and ultimately forming the PVR membrane. In this study, the signal pathways mediating cell migration of RPE induced by growth factors were investigated. Hepatocyte growth factor (HGF), epidermal growth factor (EGF) or heparin-binding epidermal growth factor (HB-EGF) induced a greater extent of migration of RPE50 and ARPE19 cells, compared with other growth factors. According to inhibitor studies, migration of RPE cells induced by each growth factor was mediated by protein kinase C (PKC) and ERK (MAPK). Moreover, HGF coupled with EGF or HB-EGF had synergistic effects on cell migration and enhanced activation of PKC and ERK, which were attributed to cross activation of growth factor receptors by heterogeneous ligands. Furthermore, using the shRNA technique, PKCδ was found to be the most important PKC isozyme involved. Finally, vitreous fluids from PVR and PDR patients with high concentration of HGF may induce RPE cell migration in PKCδ- and ERK- dependent manner. In conclusion, migration of RPE cells can be synergistically induced by HGF coupled with HB-EGF or EGF, which were mediated by enhanced PKCδ activation and ERK phosphorylation.

Activation of hepatocyte growth factor-induced apoptosis in hepatic stellate cells

To determine whether apoptosis is induced in rat hepatic stellate cells (HSCs) in response to activation of the hepatocyte growth factor (HGF) by hepatocyte growth factor activator (HGFA) by using a co-culture system of bone marrow mesenchymal stem cells (BMSCs) and HSCs. In this study, cells were divided into the following five groups: HSC control group: HSCs co-cultured with fibroblast cells; HSCs blank group: HSCs cultured alone; BMSCs blank group: BMSCs cultured alone; Experimental group: BMSCs + HSCs; HGFA intervention group: HSCs treated with 70 ng/mL of HGFA. The culture systems were established in culture plates with transwell inserts, and cells were assessed at 24, 48, and 72 h of growth. Dynamic changes in cell morphology were observed under an inverted phase contrast microscope. The surface markers of BMSCs and the apoptosis rate of HSCs were detected by Annexin-V-FITC/propidium iodide (PI). Expression of a-smooth muscle actin (SMA) in HSCs was evaluated by immunohistochemistry. The presence of activated HGF (HGF-a chain) was determined by immunofluorescent staining. HSC proliferation was measured by MTT assay, and the concentrations of HGF and HGFA were quantified by enzyme-linked immunosorbent assay (ELISA). MTT results indicated that treatment with HGF alone had no effect on HSC proliferation rate (vs. HSC blank group, P more than 0.05), but that 24 h treatment with HGFA significantly inhibited the proliferation rate (0.26 ± 0.00 vs. blank group: 0.13 ± 0.04, P = 0.02); moreover, this effect was concentration-dependent. Expression of HGF-a was lower in the experimental group than in the HGFA intervention group at 72 h (37.24 ± 1.03 vs. 40.44 ± 0.77, P = 0.04), and both of these groups had higher expression than the control group at all time points examined (P less than 0.05). The apoptosis rate was consistently higher in the experimental group than in the HGFA intervention group, but most robustly at 72 h (40.77 ± 1.16% vs. 33.35 ± 2.04%, P = 0.00); moreover, the apoptosis rate was significantly higher than that in the control group at all time points examined (P less than 0.01). The concentration of HGF in the experimental group and the HGFA intervention group showed a time-dependent reduction, and was consistently lower than that in the HSCs control group (P less than 0.05). Finally, the concentration of HGFA was higher in the experimental group than in the blank group at all time points examined (P less than 0.05). The BMSC-HSC co-culture system can promote secretion of HGFA from HSCs and HGF activation, thereby inducing apoptosis of HSCs.