Epidermal Growth Factor mRNA Research Articles

A functional polymorphism in the epidermal growth factor gene predicts hepatocellular carcinoma risk in Japanese hepatitis C patients

BackgroundA single nucleotide polymorphism (SNP) in the epidermal growth factor (EGF) gene (rs4444903) has been associated with increased risk of cancer, including hepatocellular carcinoma (HCC). The aim of this study was to examine the relationship between the EGF SNP genotype and the development and prognosis of HCC, in a Japanese population.MethodsRestriction fragment-length polymorphism was used to determine the presence of the EGF SNP genotype in 498 patients, including 208 patients with HCC. The level of EGF messenger ribonucleic acid (mRNA) expression in cancerous tissues was measured by quantitative reverse transcription polymerase chain reaction. The correlation between the EGF SNP genotype and prognosis was statistically analyzed in the patients with HCC.ResultsThe proportion of the A/A, A/G, and G/G genotypes were 5.3%, 42.8%, and 51.9%, respectively, in the patients with HCC, whereas in those without HCC, they were 8.6%, 35.9%, and 55.5%, respectively, revealing that the odds ratio (OR) of developing HCC was higher in patients with a G allele (OR =1.94, P=0.080 for A/G patients and OR =1.52, P=0.261 for G/G patients, as compared with A/A patients). In particular, when the analysis was limited to the 363 patients with hepatitis C, the OR for developing HCC was 3.54 (P=0.014) for A/G patients and was 2.85 (P=0.042) for G/G patients, as compared with A/A patients. Tumoral EGF mRNA expression in G/G patients was significantly higher than that in A/A patients (P=0.033). No statistically significant differences were observed between the EGF SNP genotype and diseasefree or overall survival.ConclusionThe EGF SNP genotype might be associated with a risk for the development of HCC in Japanese patients but not with prognosis. Of note, the association is significantly stronger in patients with hepatitis C, which is the main risk factor for HCC in Japan.

P.5.f.001 Correlations between pain intensity and cognitive dysfunction in surgery

Intrauterine growth retardation (IUGR) infants have impaired gastrointestinal function with resultant feeding difficulties and predisposition to necrotizing enterocolitis. Supplemented amniotic fluid swallowed by the developing fetus is a potential prenatal treatment for IUGR. Rabbits have naturally occurring IUGR fetuses based on uterine position. To determine intestinal response to epidermal growth factor (EGF) infusion, this rabbit model of IUGR was studied.Eight pregnant rabbits underwent placement of intraamniotic catheters into 2 normal and 2 IUGR fetuses per mother on gestational day 24 of a 31-day gestation. Miniosmotic pumps infused either EGF (about 300 μg/kg/d) or control solution forming 4 study groups (EGF-Favored [Fav] v. Cont-Fav; EGF-IUGR v. Cont-IUGR). On gestational day 31, the fetal gastrointestinal tracts were harvested for analysis. Intestinal epithelial cell proliferation was studied by 5-bromo-2-deoxyuridine (BrdU) incorporation, villus heights were measured, and EGF mRNA was measured by reverse transcriptase polymerase chain reaction (RT-PCR). Statistical analysis was performed using Students’ t test.Fetal survival rate was 87%. EGF-IUGR fetal weights were increased compared with Cont-IUGR fetuses. EGF infusion significantly increased IUGR fetal small intestinal villus height and BrdU-positive small intestinal (SI) crypt cells, all approaching Cont-Fav levels. EGF mRNA was expressed throughout the gastrointestinal tract.Supplemental amniotic EGF normalizes fetal weight and intestinal proliferation in the IUGR fetal rabbit. The inclusion of EGF in supplemental amniotic feeding solutions is supported.

Effect of mobilization bone marrow stem cells on hypoxia inducible factor-1α and epidermal growth factor in rats with renal injury induced by ischemic reperfusion

Objective To investigate the mobilization of granulocyte-colony stimulating factor(G-CSF) with stem cell factor(SCF) in bone marrow stem cells(BMSC), and to observe the therapeutic effects of BMSC mobilization on repairing ischemia/reperfusion(I/R) renal injury induced by hypoxia inducible factor-1α(HIF-1α) and epidermal growth factor(EGF), and to investigate the mechanism for BMSC mobilization for repairing I/R renal injury. Methods One hundred and sixty male Sprague-Dawley rats(age: 8-10 weeks) were randomly allocated into 4 groups(40 rats in each group): control group(group A), I/R group(group B), SCF+ G-CSF treatment group(group C) and SCF+ G-CSF control group(group D). The expressions of CD34+ and EGF were measured by immunohistochemistry technique, and the expression of EGF mRNA was measured by reverse transcriptase-polymerase chain reaction. Results 1.After I/R injury, the renal tubular epithelial cells were in a state of degeneration, necrosis, even exfoliation.As the time prolonged, the necrotic renal tubules were repaired gradually, but the repair in group C was not good compared with that in group B. The level of group C was obviously lower than that of group B on the 5th, 10th, 17th and 24th day(P 0.05). On 5 d postoperative, the level of CD34+ cells of group B and group C was conspicuous higher than group A and D(P<0.05), and that of group C was conspicuous higher than that of group B(P<0.05). They descended gradually from the 5th day with the time prolonged.3.The expressions of HIF-1α of group B and group C had a highly positive reaction on the 5th day postoperatively and the level of HIF-1α of group B and group C decended to the normal level with the time prolonged, and at each time, the expression of HIF-1α of group C was significantly higher than that of other groups(P<0.05).4.On the 5th day postoperative, the expression of EGF of group B and group C showed a higher positive reaction on the 5th day postoperatively and the level of EGF in group B and group C descended to the normal level with the time prolonged, and at each time the expression of EGF of group C was significantly higher than that of the other groups(P<0.05).5.The expressions of EGF mRNA of group B and C showed higher positive reaction on the 5th day postoperatively(P<0.05). The level of EGF mRNA of group B was higher significantly than that of group A on the 17th day posto-peratively(P<0.05). The level of EGF mRNA of group C was a peak and higher than that of other groups on the 10th day postoperatively(P<0.05). The level of EGF of group B and group C descended to a normal level with the time prolonged.6.There was a positive correlation between the expressions of HIF-1α and EGF(r=0.815, P<0.01). Conclusions Combination of SCF with the mobilization of G-CSF sufficiently homed BMSC to injured renal tissue was an important factor for the recovery of injured renal tubules.Combination of SCF with G-CSF may promote the expression of HIF-1α and EGF in the kidney may be one of the mechanisms of why combination of SCF with G-CSF can promote recovery from renal ischemic reperfusion injury. Key words: Bone marrow stem cell; Ischemic reperfusion, renal; Hypoxia inducible factor-1α; Epidermal growth factor; Rat

Nephroprotective effects of Isaria felina in rats with adenine-induced chronic renal failure

Chronic renal failure (CRF) is a progressive, life-threatening condition with limited treatment options. Cordyceps sinensis is a fungus that has nephroprotective effects, and Isaria felina (IF) is a fungus isolated from C. sinensis fruiting bodies. We evaluated IF efficacy using an adenine-induced CRF animal model. Forty male Sprague-Dawley rats were divided into normal control (n = 8) and adenine groups (n = 32; 100 mg/kg for 30 days). The adenine group was subdivided into a model control group (n = 7), a positive control group (200 mg/kg Jinshuibao capsule (JSB; n = 8), and two IF groups (200 mg/kg, n = 8; 100 mg/kg, n = 8). After treatment for 30 days, animals were narcotized and abdominal aortic blood was analysed. Kidney functions were evaluated. Higher serum creatinine, blood urea nitrogen and uric acid levels, and lower creatinine clearance was observed in the model control group compared with JSB and IF groups (P < 0.05). Red blood cell count, haemoglobin and haematocrit levels in the 200 mg/kg IF group were higher than in the model control group (P < 0.05). Transforming growth factor-β1 mRNA expression in the model control group was higher than the normal control and 200 mg/kg IF groups (P < 0.05). Epidermal growth factor mRNA in the model control group was lower than in the normal control and both IF-treated groups (P < 0.05). Structural renal damage was observed in all adenine-treated rats, but was less severe in the JSB and IF groups. IF may reverse the damaged kidney functions-induced with adenine in rats.

Development of a novel therapeutic approach using a retinoic acid-loaded microneedle patch for seborrheic keratosis treatment and safety study in humans

Seborrheic keratosis is one of the most common skin benign tumors in humans with a high occurrence rate of 80%–100% in people >50years of age; however, its pathogenesis is still unclear. The standard treatment includes cryotherapy and laser surgery for physically removing lesions. Drug therapy for this condition has not been well established. We aimed to evaluate the use of all-trans retinoic acid (ATRA)-loaded microneedle (MN) patches as a simple, alternative therapeutic option to traditional surgical treatments. This therapeutic strategy was designed to induce the proliferation of basal keratinocytes and accelerate stratum corneum turnover, leading to the lesion falling off the surface of the skin. The MN patch induced epidermal hyperplasia and marked expression of heparin-binding epidermal growth factor-like growth factor mRNA and protein corresponding to ATRA activity in the skin of HR-1 hairless mice. The acceleration of stratum corneum turnover was also observed by the dansyl chloride method. The skin irritation study in mice and safety study in humans support the safety findings of our study. Overall, MN patches can offer an effective and safe means of ATRA delivery into the skin, and the ATRA-loaded MN patch appears to be an effective pharmaceutical product providing a novel therapeutic option for seborrheic keratosis.

Neurobehavioral deficits of epidermal growth factor-overexpressing transgenic mice: Impact on dopamine metabolism

Epidermal growth factor (EGF) and its family member neuregulin-1 are implicated in the etiology of schizophrenia. Our recent pharmacological studies indicate that EGF injections to neonatal and adult rats both induce neurobehavioral deficits relevant to schizophrenia. We, however, did not evaluate the genetic impact of EGF transgene on neurobehavioral traits. Here we analyzed transgenic mice carrying the transgene of mouse EGF cDNA. As compared to control littermates, heterozygous EGF transgenic mice had an increase in EGF mRNA levels and showed significant decreases in prepulse inhibition and context-dependent fear learning, but there were no changes in locomotor behaviors and sound startle responses. In addition, these transgenic mice exhibited higher behavioral sensitivity to the repeated cocaine injections. There were neurochemical alterations in metabolic enzymes of dopamine (i.e., tyrosine hydroxylase, dopa decarboxylase, catechol-O-methyl transferase) and monoamine contents in various brain regions of the EGF transgenic mice, but there were no apparent neuropathological signs in the brain. The present findings rule out the indirect influence of anti-EGF antibody production on the reported behavioral deficits of EGF-injected mice. These results support the argument that aberrant hyper-signals of EGF have significant impact on mouse behavioral traits and dopamine metabolism.

Serum EGF and NGF levels of patients with brain injury and limb fracture

ObjectiveTo explore the expression and significance of human epidermal growth factor (EGF) and nerve growth factor (NGF) in patients with knee osteoarthritis. MethodsRT-PCR and enzyme-linked immunosorbent assay were used to measure the serum EGF and NGF expression levels of patients with limb fracture and brain trauma injurry after 1 d, 3 d, 7 d, 14 d and the relationship between them was analyzed. The level was compared among the simple fracture group, traumatic brain injury group and the normal control group, with 40 cases in each group. ResultsThe serum NGF levels were significantly different among three groups. Serum NGF, EGF mRNA and protein levels gradually decreased with the increasing injury time in the limb fracture combined with brain injury group, traumatic brain injury group, the simple fracture group and the health control group (P<0.05). ConclusionsThe serum of NGF, EGF levels significantly increased when limb fracture combined with brain injury, so EGF and NGF may be involved in the process of fracture healing.

Effects of radiation on the epidermal growth factor receptor pathway in the heart

Purpose: Radiation-induced heart disease (RIHD) is a serious side-effect of thoracic radiotherapy. The epidermal growth factor receptor (EGFR) pathway is essential for the function and survival of cardiomyocytes. Hence, agents that target the EGFR pathway are cardiotoxic. Tocotrienols protect from radiation injury, but may also enhance the therapeutic effects of EGFR pathway inhibitors in cancer treatment. This study investigated the effects of local irradiation on the EGFR pathway in the heart and tests whether tocotrienols may modify radiation-induced changes in this pathway.Methods: Male Sprague-Dawley rats received image-guided localized heart irradiation with 21 Gy. Twenty four hours before irradiation, rats received a single dose of tocotrienol-enriched formulation or vehicle by oral gavage. At time points from 2 h to 9 months after irradiation, left ventricular expression of EGFR pathway mediators was studied.Results: Irradiation caused a decrease in the expression of epidermal growth factor (EGF) and neuregulin-1 (Nrg-1) mRNA from 6 h up to 10 weeks, followed by an upregulation of these ligands and the receptor erythroblastic leukemia viral oncogene homolog (ErbB)4 at 6 months. In addition, the upregulation of Nrg-1 was statistically significant up to 9 months after irradiation. A long-term upregulation of ErbB2 protein did not coincide with changes in transcription or post-translational interaction with the chaperone heat shock protein 90 (HSP90). Pretreatment with tocotrienols prevented radiation-induced changes at 2 weeks.Conclusions: Local heart irradiation causes long-term changes in the EGFR pathway. Studies have to address how radiation may interact with cardiotoxic effects of EGFR inhibitors.

Abstract 3145: Investigation of a functional polymorphism in the epidermal growth factor gene for pathogenesis and prognosis of hepatocellular carcinoma in Japanese patients.

Abstract [Introduction] A single nucleotide polymorphism (SNP) in the epidermal growth factor (EGF) gene (rs4444903) has been investigated as a predictor of hepatocellular carcinoma (HCC). Previous reports identified an A to G transition at position 61 in the 5’ untranslated region of the EGF gene and demonstrated that the G/G genotype was associated with an increased risk of developing HCC as compared to the A/G and A/A genotype. Racial differences with respect to the distribution of this SNP have been reported; however, no investigation has examined a Japanese population. [Objectives] To examine the relationship between EGF SNP genotype and the development and prognosis of HCC in a Japanese population.[Methods] Restriction fragment-length polymorphism was used to determine EGF SNP genotype in two hundred and eight patients with HCC, who underwent resection in our department between 1994 and 2010. The level of EGF mRNA expression in cancerous tissues was measured by quantitative reverse transcription-PCR. EGF SNP was also analyzed in three hundred and four patients with either hepatitis, liver cirrhosis, or healthy controls. The correlation between EGF SNP genotype and prognosis was statistically analyzed in patients with HCC. [Results] A/A, A/G, and G/G genotype were 5.3%, 42.8%, and 51.9% in the Japanese patients with HCC. Tumoral EGF mRNA expression in G/G patients was significantly higher than that in A/A patients (P=0.04). A/A, A/G, and G/G genotype were 8.2%, 35.9%, and 55.9% in patients without HCC. Analysis of the distribution of allelic frequencies revealed that the odds ratios of developing HCC were 1.86 (95% CI 0.88-4.12, P=0.23) in A/G patients and 1.44 (95% CI 0.70-3.17, P=0.33) in G/G patients compared with A/A patients. However, when the analysis was limited to patients with hepatitis C, the odds ratio were 3.37 (95% CI 1.20-12.06, P=0.02) in A/G patients, and 2.77 (95% CI 1.10-9.78, P=0.04) in G/G patients, compared with A/A patients. 3) No statistically significant differences were observed between EGF SNP genotype and disease-free and overall survival. [Conclusions] The frequency of the EGF SNP genotype in a Japanese population was characterized by a very low distribution of A/A patients. The EGF 61*G allele was associated with greater EGF mRNA expression in HCC tissue. The EGF SNP genotype was also associated with risk for development of HCC in hepatitis C patients but not with prognosis. Citation Format: Masaya Suenaga, Suguru Yamada, Tsutomu Fujii, Bryan C. Fuchs, Norio Okumura, Goro Nakayama, Shin Takeda, Kenneth K. Tanabe, Yasuhiro Kodera. Investigation of a functional polymorphism in the epidermal growth factor gene for pathogenesis and prognosis of hepatocellular carcinoma in Japanese patients. [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 3145. doi:10.1158/1538-7445.AM2013-3145

Glucose transporter member 1 is involved in UVB‐induced epidermal hyperplasia by enhancing proliferation in epidermal keratinocytes

Glucose transporter member 1 (GLUT-1) is one of the major facilitated glucose transporters and contributes to the promotion of keratinocyte proliferation in psoriasis and carcinogenic lesions. In this study, we postulate that GLUT-1 is involved in ultraviolet B (UVB)-induced epidermal hyperplasia. The purpose of this study is to investigate the possible role of GLUT-1 in UVB-induced hyperplasia. The effects of UVB on GLUT-1 expression levels were investigated in in vitro and in vivo studies. In addition, the involvement of epidermal growth factor (EGF) and hypoxia inducible factor-1 alpha (HIF-1α), transcriptional factors for GLUT-1, in GLUT-1-related events were investigated. GLUT-1 mRNA and its protein levels were markedly increased by UVB irradiation in HaCaT cells. In in vivo studies, a strong immunofluorescence signal of GLUT-1 was clearly observed around the basal layer of the epidermis, which proliferated excessively by UVB irradiation. In HaCaT cells, EGF mRNA and its protein levels were markedly increased by UVB irradiation, and then the GLUT-1 mRNA level was significantly increased by treatment with EGF. Additionally, the upregulation of GLUT-1 by both UVB irradiation and treatment with EGF was significantly suppressed by transfection with HIF-1α siRNA. We conclude that GLUT-1 is involved in UVB-induced epidermal hyperplasia by enhancing proliferation of epidermal basal cells, and the GLUT-1-related event might be regulated by an increase in HIF-1α stimulated by EGF.

The effects of epidermal growth factor (EGF) on the in vitro development of isolated goat secondary follicles and the relative mRNA expression of EGF, EGF-R, FSH-R and P450 aromatase in cultured follicles

The effects of varying concentrations of EGF were evaluated in terms of in vitro follicular development and the mRNA expression levels of EGF, EGF-R, FSH-R and P450 aromatase. After 6days, the addition of 50ng/mL of EGF to the culture medium increased the antrum formation rates in comparison to cultured control and after 18days of culture produced oocytes with higher rates of meiosis resumption when compared to the other treatments (P<0.05). The daily follicular growth rates in presence of EGF (50 or 100) were increased in comparison to the cultured control (P<0.05). Treatment with EGF 50 stimulated the expression of EGF mRNA but reduced EGF-R mRNA expression and estradiol secretion as compared to the cultured control (P<0.05). After 18days of culture, the mRNA levels for FSH-R and P450 aromatase were greater than those of the non-cultured controls (P<0.05). In conclusion, the effects of EGF treatment on the mRNA levels for EGF, EGF-R, FSH-R, and P450 aromatase varied according to the stage of follicle development.

Substance P affects growth factors in Pseudomonas aeruginosa-infected mouse cornea.

This study analyzed the influence of substance P (SP) on growth factors related to wound healing in mice in the presence of infectious keratitis. Naturally resistant mice were injected intraperitoneally with SP or phosphate-buffered saline and infected with Pseudomonas aeruginosa, and corneal messenger RNA (mRNA) levels of growth factors and apoptosis genes were tested. Enzyme-linked immunosorbent assay determined the protein levels, whereas immunohistochemistry tested the distribution, macrophage phenotype, and cell quantitation. In vitro, macrophages were stimulated with lipopolysaccharide (LPS; with or without SP) and mRNA levels of proinflammatory and antiinflammatory cytokines and apoptosis genes were tested. After SP, epidermal growth factor mRNA and protein levels were disparately regulated early, with no differences later in the disease. Hepatocyte growth factor and fibroblast growth factor-7 mRNA and protein levels were increased after SP treatment. Enumerating dual-labeled stromal cells revealed no difference between SP-treated versus phosphate-buffered saline-treated groups in the percentage of epidermal growth factor-labeled fibroblasts or macrophages, but there were significant increases in both hepatocyte growth factor- and fibroblast growth factor-7-labeled cells. Type 2 (M2) macrophages and caspase-3 mRNA levels were decreased, whereas B-cell lymphoma-2 mRNA expression was increased after SP treatment. In vitro, mRNA levels of several proinflammatory cytokines and B-cell lymphoma-2 were elevated, whereas transforming growth factor β was decreased after macrophage stimulation with SP (with LPS) over LPS alone. (Mice: n = 105 control; 105 experimental.) These data show that treatment with SP in infectious keratitis elevates growth factors but also adversely affects the disease by enhancing the inflammatory response and its sequelae.

742P - Investigation of a Functional Polymorphism in the Epidermal Growth Factor Gene for Pathogenesis and Prognosis of Hepatocellular Carcinoma in Japanese Patients

ABSTRACT Introduction A single nucleotide polymorphism (SNP) in the epidermal growth factor (EGF) has been investigated as a predictor of hepatocellular carcinoma (HCC). Previous reports demonstrated that an A to G transition at position 61 in the 5′ untranslated region of the EGF gene was identified, and that G/G genotype was associated with an increased risk of developing HCC compared with the A/G and A/A genotype. It has been reported that there are differences in the distribution of polymorphisms by race, there is no report in the Japanese population. Objectives To examine the relationship between EGF gene polymorphism genotype and the development and prognosis of HCC in the Japanese population. Methods Restriction fragment-length polymorphism was used to determine the EGF gene polymorphism genotype in two hundred and thirteen patients with HCC, who underwent resection in our department between 1996 and 2010. The correlation between these values and clinicopathological factors and prognosis were statistically analyzed. 2) The level of EGF mRNA expression in cancerous tissues was measured by quantitative reverse transcription-PCR. 3) EGF SNP was also analyzed in one hundred and forty patients with hepatitis and liver cirrhosis. Results 1) A/A, A/G, and G/G genotype were 5.2%, 43.7%, and 51.2% in the Japanese patients with HCC. A/A patients had better overall and disease-free prognosis than other genotypes, although no significant differences were found. 2) EGF mRNA expression in G/G patients was significantly higher than that in A/A patients (P = 0.04). 3) A/A, A/G, and G/G genotype were 7.1%, 34.3%, and 58.6% in patients without HCC. Analysis of the distribution of allelic frequencies revealed that odds ratio of developing HCC was 1.76 (95% CI 0.69-4.47, P = 0.23) in A/G patients, and 1.21 (95% CI 0.48-3.00, P = 0.68) in G/G patients, compared with A/A patients. Conclusions EGF 61*G allele was associated with greater transcript stability and EGF expression in the Japanese population. The EGF gene polymorphism genotype was associated with risk for development of HCC. Disclosure All authors have declared no conflicts of interest.

Deletion of the epidermal growth factor receptor in renal proximal tubule epithelial cells delays recovery from acute kidney injury

To determine the role of epidermal growth factor receptor (EGFR) activation in renal functional and structural recovery from acute kidney injury (AKI), we generated mice with a specific EGFR deletion in the renal proximal tubule (EGFRptKO). Ischemia–reperfusion injury markedly activated EGFR in control littermate mice; however, this was inhibited in either the knockout or wild-type mice given erlotinib, a specific EGFR tyrosine kinase inhibitor. Blood urea nitrogen and serum creatinine increased to a comparable level in EGFRptKO and control mice 24 h after reperfusion, but the subsequent rate of renal function recovery was markedly slowed in the knockout mice. Twenty-four hours after reperfusion, both the knockout and the inhibitor-treated mice had a similar degree of histologic renal injury as control mice, but at day 6 there was minimal evidence of injury in the control mice while both EGFRptKO and erlotinib-treated mice still had persistent proximal tubule dilation, epithelial simplification, and cast formation. Additionally, renal cell proliferation was delayed due to decreased ERK and Akt signaling. Thus, our studies provide both genetic and pharmacologic evidence that proximal tubule EGFR activation plays an important role in the recovery phase after acute kidney injury.

The TAM-family receptor Mer mediates production of HGF through the RhoA-dependent pathway in response to apoptotic cells

The TAM receptor protein tyrosine kinases Tyro3, Axl, and Mer play important roles in macrophage function. We investigated the roles of the TAM receptors in mediating the induction of hepatocyte growth factor (HGF) during the interaction of macrophages with apoptotic cells. Mer-specific neutralizing antibody, small interfering RNA (siRNA), and a recombinant Mer protein (Mer/Fc) inhibited HGF mRNA and protein expression, as well as activation of RhoA, Akt, and specific mitogen-activated protein (MAP) kinases in response to apoptotic cells. Inhibition of Axl or Tyro3 with specific antibodies, siRNA, or Fc-fusion proteins did not prevent apoptotic cell-induced HGF mRNA and protein expression and did not inhibit activation of the postreceptor signaling molecules RhoA and certain MAP kinases, including extracellular signal-regulated protein kinase and c-Jun NH(2)-terminal kinase. However, Axl- and Tyro3-specific blockers did inhibit the activation of Akt and p38 MAP kinase in response to apoptotic cells. In addition, none of the TAM receptors mediated the effects of apoptotic cells on transforming growth factor-β or epidermal growth factor mRNA expression. However, they were involved in the induction of vascular endothelial growth factor mRNA expression. Our data provide evidence that when macrophages interact with apoptotic cells, only Mer of the TAM-family receptors is responsible for mediating transcriptional HGF production through a RhoA-dependent pathway.

RNA interference-mediated downregulation of hypoxia-inducible factor-1α inhibits angiogenesis and survival of oral squamous cell carcinoma in vitro and in vivo

Hypoxia-inducible factor-α (HIF-1α) is the key transcription factor involved in the adaptive response to hypoxia. It has been established that HIF-1α is overexpressed in various human cancers, including oral squamous cell carcinoma (OSCC), and orchestrates a wide spectrum of many key cancer molecular pathways involved in diverse tumor progression. In this study, RNA interference (RNAi) was introduced to downregulate the expression of HIF-1α in OSCC in vitro and in vivo. The mRNA and protein expression levels of HIF-1α and vascular epidermal growth factor were detected by quantitative real-time reverse transcription-PCR, western blot, and immunohistochemistry, and cell proliferation activity was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Tumor growth, microvascular density, apoptosis, and the expression of proliferation-related gene Ki-67 in tumor xenografts were evaluated. Our data demonstrated that small interfering RNA targeting HIF-1α significantly inhibited the mRNA and protein expression levels of HIF-1α and vascular epidermal growth factor. In addition, marked suppression of tumor cell prolifeation and xenograft growth, significant microvascular density suppression, reduced expression of Ki-67, and increased cell apoptosis were observed. These findings suggest that RNAi targeting HIF-1α could effectively inhibit the progression of OSCC. Thus, it may be used as a potent and specific therapy for oral cancer, especially in inhibiting and preventing cancer cell angiogenesis and survival.

Local Expression of Epidermal Growth Factor–Like Growth Factors in Human Testis and Its Role in Spermatogenesis

Microdissection testicular sperm extraction (micro-TESE) has revealed that spermatogenesis in nonobstructive azoospermia (NOA) patients is heterogeneous, even in the same testis, but there is no information regarding growth factors to support spermatogenesis. We investigated the involvement of epidermal growth factor (EGF)-like growth factors, which play important roles in cell proliferation and differentiation in NOA patients who underwent micro-TESE. Testicular samples were obtained from 5 fertile men (15 samples), 5 prostate cancer patients receiving maximum androgen blockade (10 samples), and 13 NOA patients who underwent micro-TESE (50 samples). The expression of the mRNA for EGF, heparin binding (HB)-EGF, amphiregulin, epiregulin, betacellulin, and transforming growth factor (TGF)-α were analyzed by real-time polymerase chain reaction analysis and adjusted using the expression of glyceraldehyde-3-phosphate dehydrogenase. Heterogeneous expression of these EGF-like growth factors were observed even in the same testis. The expression of HB-EGF, amphiregulin and TGF-α in NOA and prostate cancer patients was significantly lower than observed in fertile controls. In NOA patients, expression in the testicular sample comprising mature sperm was significantly higher than those without mature sperm, indicating that HB-EGF, amphiregulin, and TGF-α are considered to participate in creating a suitable niche for spermatogenesis. Considering the findings that ablation of gonadotropin inhibited and human chorionic gonadotropin stimulation increased these EGF-like growth factors, the expressions are presumably under gonadotropin regulation.

Runx3 Expression and Its Roles in Mouse Endometrial Cells

Runx3 is a transcription factor that belongs to the Runx family. We studied the localization of Runx3 mRNA in the mouse uterus, and its function in the mouse endometrium using Runx3 knockout (Runx3(-/-)) mice. Runx3 mRNA was detected in the endometrial luminal epithelial cells, glandular epithelial cells and stromal cells below the epithelial cell layer on the luminal side. The uteri of Runx3(-/-) mice were smaller than those of wt mice. The endometrial layer and uterine glands of Runx3(-/-) mice were less developed than those of wild-type mice, and the endometrial stromal layer was thinner. Transforming growth factor β1 and β3 (TGFβ1 and β3) mRNA levels in endometrial stromal cells of Runx3(-/-) mice were low compared with those of wild-type mice. Estradiol-17β (E2) increased Tgfb2 mRNA levels in endometrial stromal cells of Runx3(-/-) mice, but not in those of wild-type mice. E2 increased epidermal growth factor (EGF) mRNA levels in endometrial stromal cells of wild-type mice, but did not increase those of Runx3(-/-) mice. The diminished Tgfb1 and Tgfb3 mRNA expressions may lead to the reduced proliferation of endometrial stromal cells. Alterations of E2-associated expressions of Tgfb2 and Egf mRNA in endometrial stromal cells of Runx3(-/-) mice may be associated with suppression of E2-dependent endometrial epithelial cell proliferation in Runx3(-/-) mice. Thus, Runx3 is likely to be a regulatory factor responsible for endometrial growth.

Goat Oocyte Production by Standard or One‐shot FSH Treatments and Quantitative Analysis of Transcripts for EGF Ligands and its Receptor after In Vitro Maturation

Hormonal ovarian stimulation may affect the success of embryo production by regulating transcripts in recovered cumulus-oocyte complexes (COCs). Here, in parallel to morphological classification and in vitro maturation (IVM) rate analysis, we investigated the expression of epidermal growth factor (EGF) and its receptor (EGFR) in oocytes and cumulus cells from goat COCs recovered by laparoscopy after standard [multi-dose follicle-stimulating hormone (FSH)] or one-shot (single dose FSH plus eCG) treatments. No differences were observed among the number of recovered and morphologically graded COCs or the IVM rates for both gonadotropic treatments. However, the standard protocol produced COCs with higher EGFR expression in the cumulus cells than the one-shot treatment. Additionally, EGF mRNA levels were less than EGFR mRNA levels, and they did not differ among COCs from both treatments. However, during maturation, the EGF transcripts increased in oocytes derived only from the standard protocol. Interestingly, IVM strikingly increased EGFR expression in oocytes and cumulus cells but not in oocytes that fail in first polar body extrusion, irrespective of hormonal treatment. These results appear to be related to the resumption of meiosis and suggest that EGF may act through the cumulus cells or directly on the oocyte receptor.

Postnatal regression of hypertrophic cardiomyopathy in infants of diabetic mothers: A crosstalk between Hox genes and epidermal growth factor (EGF) gene polymorphism?

mothers: A crosstalk between Hox genes and epidermal growth factor (EGF) gene polymorphism? Raffaella Mormile ⁎, Giorgio Vittori , Mario De Michele , Umberto Squarcia , Federico Quaini e a Division of Pediatric and Neonatology, Moscati Hospital, Aversa, Italy b Division of Gynecology, San Carlo di Nancy Hospital, Rome, Italy c Division of Cardiology, Moscati Hospital, Aversa, Italy d Pediatric Cardiology, University of Parma, Italy e Department of Internal Medicine and Biomedical Sciences, Section of Internal Medicine, University of Parma, Italy