Pro-and Anti-inflammatory Biomarkers as Potential Prognosis Markers in Oral Squamous Cell Carcinoma

ABSTRACT: The most widely used method to classify prognostic factors in cancers today is TNM. However, Oral Squamous Cell Carcinoma (OSCC) often demonstrates different behaviors in relation to aggressiveness and therapeutic response at the same TNM stage. So, in such cases biomarkers can be used to identify the biological diversity of these tumors more reliably, leading to better therapeutic strategies and disease management. The presence of inflammatory immune cells in the tumor microenvironment can have pro or antitumor effects and the investigation of the expression of inflammatory markers in OSSC can be usefulto design immunotherapeutic interventions. The Transforming Growth Factor alpha is a potent stimulator of cell migration that acts on cell proliferation, invasion and metastasis of cancer, as well as immune suppression and angiogenesis. Inflammatory cytokines, such as Interferon-gamma, mediate macrophage differentiation. Macrophages are an important component of the OSCC microenvironment. The greater amount of tumor-associated macrophages, especially the M2 phenotype, may be associated with a more aggressive biological behavior of the OSCC and, consequently, with reduced survival.

RESUMEN: El método más utilizado para clasificar los factores de pronóstico en los cánceres en la actualidad es TNM. Sin embargo, el carcinoma oral de células escamosas (COCE) a menudo muestra diferentes comportamientos en relación con la agresividad y la respuesta terapéutica en la misma etapa TNM. Entonces, en tales casos, los biomarcadores pueden usarse para identificar la diversidad biológica de estos tumores de manera más confiable, lo que lleva a mejores estrategias terapéuticas y manejo de la enfermedad. La presencia de células inmunes inflamatorias en el microambiente tumoral puede tener efectos pro o antitumorales y la investigación de la expresión de marcadores inflamatorios en COCE puede ser útil para diseñar intervenciones inmunoterapéuticas. El factor de crecimiento transformante α es un potente estimulador de la migración celular que actúa sobre la proliferación celular, la invasión y metástasis del cáncer, así como la inmunosupresión y la angiogénesis. Las citocinas inflamatorias, como el IFN-γ, median en la diferenciación de macrófagos. Los macrófagos son un componente importante del microambiente COCE. La mayor cantidad de macrófagos asociados a tumores, especialmente el fenotipo M2, puede estar asociada a un comportamiento biológico más agresivo del COCE y, en consecuencia, a una menor supervivencia.

DHEA and frontal fibrosing alopecia: molecular and physiopathological mechanisms

Abstract The transforming growth factor-beta 1 (TGFβ1) promotes fibrosis, differentiating epithelial cells and quiescent fibroblasts into myofibroblasts and increasing expression of extracellular matrix. Recent investigations have shown that PPAR (peroxisome proliferator-activated receptor*) is a negative regulator of fibrotic events induced by TGFβ1. Dehydroepiandrosterone (DHEA) is an immunomodulatory hormone essential for PPAR functions, and is reduced in some processes characterized by fibrosis. Although scarring alopecia characteristically develops in the female biological period in which occurs decreased production of DHEA, there are no data in the literature relating its reduction to fibrogenic process of this condition. This article aims to review the fibrogenic activity of TGFβ1, its control by PPAR and its relation with DHEA in the frontal fibrosing alopecia.

Menetrier's Disease: Its Mimickers and Pathogenesis

Menetrier's disease is a rare protein-losing hypertrophic gastropathy. Histologically, it can be mistaken for other disorders showing hypertrophic gastropathy. The pathogenesis of Menetrier's disease is not fully understood; however, it appears that the epidermal growth factor receptor (EGFR) ligand, transforming growth factor alpha, contributes to the pathogenesis of this disorder. In this review, we will discuss disease entities that can mimic Menetrier's disease and the role of EGFR signaling in Menetrier's disease.

Salivary transforming growth factor alpha in patients with Sjögren's syndrome and reflux laryngitis / Estudo da concentração salivar do fator transformador de crescimento alfa em indivíduos com Sindrome de Sjögren e refluxo laringofaríngeo

Introduction: Saliva plays a key role in the homeostasis of the digestive tract, through its inorganic components and its protein growth factors. Sjögren's syndrome patients have a higher prevalence of gastroesophageal reflux disease and laryngopharyngeal reflux. Decreased salivary transforming growth factor alpha levels were observed in dyspeptic patients, but there have been no studies in patients with Sjögren's syndrome and laryngopharyngeal reflux. Objective: To compare the salivary transforming growth factor alpha levels of patients with Sjögren's syndrome and laryngopharyngeal reflux to those of healthy controls. Methods: This is a prospective controlled study. Twelve patients with Sjögren's syndrome and laryngopharyngeal reflux and 11 controls were prospectively evaluated. Spontaneous and stimulated saliva samples were obtained to establish salivary transforming growth factor alpha concentrations. Results: The salivary transforming growth factor alpha levels of patients were significantly higher than those of healthy controls. Five patients with laryngopharyngeal reflux also had erosive esophagitis; their salivary transforming growth factor alpha levels were comparable to controls. Conclusion: Salivary transforming growth factor alpha level was significantly higher in patients with Sjögren's syndrome and laryngopharyngeal reflux when compared to the control group. .

Introdução: A saliva exerce influência primordial na homeostase do sistema digestório, pelos seus componentes inorgânicos e pelos fatores de crescimento. Indivíduos com sindrome de Sjögren (SS) apresentam maior incidência da doença do refluxo gastroesofágico (DRGE) e do refluxo laringofaríngeo (RLF). Concentrações salivares diminuídas do fator transformador de crescimento alfa (TGF-α) foram observadas em doentes dispépticos, porém não há estudos em populações com SS e RLF. Objetivo: Comparar concentrações salivares do TGF-α; de indivíduos com SS e RLF a de controles saudáveis. Método: Trata-se de um estudo prospectivo controlado. Doze pacientes com SS e RLF e 11 indivíduos controles saudáveis tiveram amostras salivares espontâneas e estimuladas coletadas para estabelecer concentração de TGF-α. Resultados: A concentração salivar de TGF-α; foi estatisticamente maior no grupo estudo para ambas amostras. Este aumento foi confirmado nos sete indivíduos do grupo estudo que não apresentavam esofagite erosiva quando comparados ao grupo controle, porém não houve diferença estatística da concentração de TGF-α; entre pacientes do grupo estudo que apresentava mesofagite erosiva em comparação ao grupo controle. Conclusão: A concentração salivar de TGF-α; foi estatisticamente maior no grupo de indivíduos com SS e RLF, sem esofagite erosiva. .

Bone remodeling at microscrew interface near extraction site in the beagle dog mandible-histologic and immunohistochemical analyses

Extraction is often used as part of orthodontic therapy, and good control of anchorage is a key step after extraction. Although microscrews can be implanted close to the extraction site in order to achieve orthodontic support, the efficiency of bone remodeling at the implant-bone interface near the extraction region is dubious. OBJECTIVE: The purpose of this study was to investigate bone remodeling of the bone-microscrew interface near the tooth extraction site, in the absence of loading. MATERIAL AND METHODS: Third and fourth premolars were extracted from the mandibles of beagle dogs, followed by placement of test microscrews near the extraction sites. Control microscrews were placed further away from the extraction site. All samples were collected after 1, 3, 8, or 12 weeks of healing following extraction. The bone remodeling process at the interface was evaluated using histologic and immunohistochemical analyses. RESULTS: Initially, a large number of inflammatory cells were aggregated at the interface. The expression levels of core binding factor (Cbfa1), osteocalcin (OC) and transforming growth factor beta (TGF-β) were inconspicuous in both groups, whereas tumor necrosis factor alpha (TNF-α) was strongly expressed, especially in the test groups (P<0.05). Subsequently, the expression levels of Cbfa1, OC and TGF-β were found to increase significantly, and active osteogenesis was observed. CONCLUSIONS: During week 1, inflammatory reaction is a major concern at the bone-microscrew interface near the extraction site. However, with healing, the influence of extraction on the remodeling of bone surrounding the microscrews decreases, thus facilitating successful treatment. .

Effect of Toll-like receptor 2 on the proliferation of human keratinocytes / 中华皮肤科杂志

Objective To investigate the effect of Toll-like receptor 2(TLR2)on the proliferation of human keratinocytes.Methods Keratinocytes were isolated from the foreskin of children,and subjected to primary culture.Atier 3-5 passages.the kemtinocytes were incubated with various concentrations of peptidoglycan(PGN).a TLR2 agonist.Cell proliferation was detected by MTT colorimetric assay and the suitable concentrations of PGN were determined.The mRNA and protein expressions of Ki67.TLR2.NF-kB p65 and TGF-α were detected by real-time quantitative PCR and Western blot.respectively,in keratinocytes treated witll PGN of 0,1.25,2.5 and 5 μg/mL.Antibody blocking test was utilized to evaluate the effect of blocking TLR2 with specific anti-TLR2 neutralizing monoclonal antibody before incubation with PGN on the expressions of Ki67,TLR2,NF-KB p65 and TGF-α by keratinocytes.Results The proliferation of kemtinocytes was significantly promoted by the incubation with PGN of 1.25,2.5 and 5μg/mL for 24 hours (all P＜0.05),which also increased the expression of Ki67 protein,TLR2 mRNA and protein,and NF-KB p65 protein.Further more,the mRNA expression of Ki67 in keratinocytes was elevated bv PGN of 1.25 and 2.5μg/mL,the mRNA expression of NF-KB p65 elevated by PGN of 1.25μg/mL,and the expressions of TGF-αprotein and mRNA elevated by PGN of 1.25 and 5μg/mL (P＜0.05).The mRNA and protein expressions of Ki67,TLR2,NF-kB p65 and TGF-αwere all inhibited by the blocking of TLR2 before incubation with PGN (a11 P＜0.05).Conclusion Activation of TLR2 bv PGN could induce the over-proliferation of human keratinocytes,likely through promoting NF-rB activation and TGF-α expression.

Regulation of transforming growth factor alpha on glutamate transpoter of retinal M(u)ller cells in mice / 中华眼底病杂志

Objective To observe the regulation effect of transforming growth factor alpha(TGFα) on expression of glutamate transporter(GLAST)and ingestion activity of retinal M(u)ller cells in mice.Methods To take the retinal tissue of Kunming mouse at postnatal 7～10 day.and then cultured M(u)ller cells according to literature.The 3～4 generation cultured cells of the same primary cell were divided into two groups at random:①TGFα group:maintained in different concentrations of TGFα as 50,75,125 and 150 ng/ml,3 holes in each concentration;②Control group:cultured by Eagle culture medium which improved from Dulbeccon and contained 20%fetal calf serum.The influence of different concentrations TGFα on GLAST activity in M(u)ller cells were observed by L-3H-glutamate uptake detection;the expression of GLAST mRNA in M(u)ller cells was determined by RT-PCR;the expression of GLAST protein was detected with immunocytochemical staining.Results With the increase of TGFα concentration.both L-3H-glutamate uptake and GLAST mRNA expression were increased.The L-3H-glutamate accumulation had got to the maximum uptake at concentration of 125 ng/ml,which was 266% of that in control group,meanwhile,the expressions of GLAST mRNA also got to the maximum as 4 times of control group.Immunocytochemical staining indicated that the effect of 125ng/ml TGFα on expression of GLAST protein was higher than that in the control group,the differences between two groups were statistically significant(P＜0.05).Conclusion TGF-α can increase GLAST activity through up-regulating the expression of GLAST mRNA and protein.

Dynamic changes of epidermal growth factor and its receptor in neonatal rats with intestine injury / 中华围产医学杂志

Objective To determine the dynamic changes of ileum epidermal growth factor(EGF),transforming growth factor-α(TGF-α)and epithelial growth factor receptor(EGFR)in neonatal rats with intestinal injury and to investigate their role in the neonatal necrotizing enterocolitis(NEC). Methods One-day-old Wistar rat pups were divided into two groups.The rats in the lipopolysaccharide(LPS)-injected group(n=40)received an intraperitoneal injection with 5 mg/kg LPS and those in the control group(n=8)did not.Control group and LPS-injected group at 1,3,6,12 and 24 h following LPS challenge were sacrificed for histological evaluation of NEC and for measurernents of EGF,EGFR,EGFR mRNA and TGF-α mRNA. Results Compared with the control group(0.12 0.17).the LPS-iniected pups showed a significant increase in injury scores at 1,3,6,12 and 24 h(1.28±0.62,1.75±0.74,1.98±0.75,2.85±0.41 and 2.35士0.63,respectively)(P<0.01).EGF protein levels at 1,3,6,12,24 h[(235.9±44.3)pg/mg prot,(231.8±30.0)pg/mg prot,(223.3±48.1)pg/mg prot,(211.7±47.0)pg/mg prot and(221.4±39.0)pg/mgprot,respectively]were significantly lower than that of the control group[(287.7±42.6)pg/mg prot](P<0.05).There was a negative correlation between the EGF levels and the grade of intestinal injury within 24 h(r=1.000,P<0.01).The expression of EGFR protein and mRNA was increased after LPS administration.There was no correlation between the EGFR protein and the grade of intestinal injury(r=0.800,P>0.05).The expression of TGF-α mRNA was significantly up-regulated at 1 and 3 h following LPS injection. Conclusions Reduced levels of EGF may play a pivotal role in the pathogenesis of NEC.

Expression of ErbB Receptor Proteins and TGF-alpha during Diethylnitrosamine-induced Hepatocarcinogenesis in the Rat Liver / 대한간학회지

BACKGROUND/AIMS: ErbB receptor proteins are transmembrane tyrosine kinase receptors; when they are activated by interaction with ligands, they generate diverse cellular responses, especially during lesion development and progression to cancer. In this study the expression of ErbB receptors and TGF-alpha were investigated using an experimental cirrhosis rat model giving rise to hepatocellular neoplasms, similar to human liver diseases. METHODS: Fifty three male rats received intraperitoneal injection of diethylnitrosamine (DEN, 50 mg/kg), weekly for 18 weeks. Until the eighth week, two rats were sacrificed every two weeks and from the tenth to the eighteenth week, five rats were sacrificed weekly. Grossly, dyschromatic and dysmorphic nodules were counted and categorized into three groups: N1/N2/N3: 3 mm or = 10 mm in diameter. All nodules were examined, histologically. Antibodies for GSTp, TGF-alpha, EGF-R, ErbB2, ErbB3 and ErbB4 were used for immunohistochemistry. RESULTS: The onset of cirrhoses was noted from the twelfth week. Preneoplastic foci, hepatocellular adenomas (HCA) and hepatocellular carcinomas (HCC) were noted from the second, eleventh and fifteenth week, respectively. The nodules (N1/N2/N3: 397/258/64) included regenerating nodule; RN (N1/N2/N3: 72.3%/15.9%/0%), HCA (N1/N2/N3: 27.2%/82.2%/7.6%) and HCC (N1/N2/N3: 0.5%/ 1.9%/92.4%). EGF-R was expressed in 12.5% of RN, 64.7% HCA and 75.2% HCC. TGF-alpha was expressed in 92.4% of RN, 91.3% HCA and 93.2% HCC. Sixty eight percent of TGF-alpha expressing nodules showed concurrent EGF-R expression. ErbB2 was expressed in 83.6% of RN, 72.9% HCA and 88.7% HCC. ErbB4 was expressed in 95.2% of RN, 86.3% HCA and 62.5% HCC. CONCLUSIONS: Increased expression of EGF-R and decreased expression of ErbB4, might be related with tumor progression during DEN-induced hepatocarcinogenesis.

Mechanism of Study on Gastric Mucosal Lesion Restoration in Rats Treated by Electroacupuncture / 中国中医药信息杂志

Objective Through comparing the similarities and differences between respairing effect on rat’s gastric mucosal lesion by electroacupure (EA) at Meridian of Foot Yangming and Meridian of Foot Shaoyang, to explore the specificity of the correlation between Meridian of Foot and Stomach and the mechanism of the healing effect by EA at meridian of Foot Yangming for curing the gastric lesion. Methods Forty rats were randomly divided into 4 groups:blank group, model group, stomach tunnel group, gallbladder tunnel group. All rats (except normal group) were made model by water restraint stress (WRS). After corresponding experimental process, epidermal growth factor (EGF) and transforming growth factor alpha (TGF-?) of plasma, gastric mucous tissue concentration was detected with radioimmunoassay, and the expression of EGFR mRNA of above tissue was detected by the reverse transcriptase-polymease chain reaction (RT-PCR) method. Results The gastric mucosal lesion index of Foot Yangming group decreased significantly in comparison with that of other groups, while EGF, TGF-? of gastric mucous tissue concentration level of Foot Yangming group was obviously higher than that of model group and gallbladder tunnel group. EA oould up-regulate the expression gastric mucosal EGFRmRNA, and there was a more significant difference in the group by EA at Meridian of Foot Yangming than that in the model group and gallbladder tunnel group. Conclusion The curative effect of EA at points of Meridian of Foot Yangming on the gastric mucosa may be realized by the synthesis, secretion and release of the related EGF family factor, such as EGF, TGF-? and so on. Such special regulation of electroacupuncture on gastric mucosal tissue was related to the gene expression of EGFRmRNA, and that may be further mechanism of the curative effect of EA at points of Meridean of Foot-Yangming on gastric mucosal lesion. There existed corresponding correlation between Meridian of Foot Yangming and Stomach.

Effects of Neurotrophic Factors on the Generation of Functional Dopamine Secretory Neurons Derived from in vitro Differentiated Human Embryonic Stem Cells / 대한불임학회지

OBJECTIVE: This study was to examine the in vitro neural cell differentiation patterns of human embryonic stem (hES) cells following treatment of various neurotrophic factors [basic fibroblast growth factor (bFGF), retinoic acid (RA), brain derived neurotrophic factor (BDNF) and transforming growth factor (TGF)-alpha], particulary in dopaminergic neuron formation. METHODS: The hES cells were induced to differentiate by bFGF and RA. Group I) In bFGF induction method, embryoid bodies (EBs, for 4 days) derived from hES were plated onto gelatin dish, selected for 8 days in ITSFn medium and expanded at the presence of bFGF (10 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14 and 21 days. Group II) For RA induction, EBs were exposed of RA (10-6 M) for 4 days and allowed to differentiate in N2 medium for 7, 14 and 21 days. Group III) To examine the effects of additional neurotrophic factors, bFGF or RA induced cells were exposed to either BDNF (10 ng/ml) or TGF-alpha (10 ng/ml) during the 21 days of final differentiation. Neuron differentiation and dopamine secretion were examined by indirect immunocytochemistry and HPLC, respectively. RESULTS: The bFGF or RA treated hES cells were resulted in similar neural cell differentiation patterns at the terminal differentiation stage, specifically, 75% neurons and 11% glial cells. Additionally, treatment of hES cells with BDNF or TGF-alpha during the terminal differentiation stage led to significantly increased tyrosine hydroxylase (TH) expression of a dopaminergic neuron marker, compared to control (p<0.05). In contrast, no effect was observed on the rate of mature neuron (NF-200) or glutamic acid decarboxylase-positive neurons. Immunocytochemistry and HPLC analyses revealed the higher levels of TH expression (20.3%) and dopamine secretion (265.5+/-62.8 pmol/mg) in bFGF and TGF-alpha sequentially treated hES cells than those in RA or BDNF treated hES cells. CONCLUSION: These results indicate that the generation of dopamine secretory neurons from in vitro differentiated hES cells can be improved by TGF-alpha addition in the bFGF induction protocol.

To amplify and determine the DNA fragment of the human transforming growth factor alpha (hTGF-A) gen encoding from placental sample

The DNA fragment of the human transforming growth factor alpha gene encoding for the mature hTGF-A peptide containing 50 amino acids was amlified from placental total RNA. The nucleotid sequence obtained was continue analyzed by DNA club software. The results: they have transformed a DNA fragment (from nucleotid position 149 to 289) of TGF-A gene from Vietnamese placental total RNA

Postnatal Development of Transforming Growth factor-alpha-immunoreactive Neurons in the Diencephalon of the Rat / 체질인류학회지

Transforming growth factor-alpha (TGF-alpha) immunoreactivity during postnatal development was examined in the rat diencephalon using immunohistochemistry. The time of appearance and localization of TGF-alpha immunoreactivity was slightly different in many areas of diencephalon during postnatal development. At birth, TGF-alpha immunoreactivity was mainly evident in thalamic medial, median and parafascicular thalamic nucleus of intralaminar nuclei. In addition, TGF-alpha immunoreactivity was clearly evident at the first postnatal week in most hypothalamic nuclei. Therefore, TGF-alpha immunoreactivity was found at postnatal days 7 in most diencephalic nuclei excepting the vental posterior thalamic nuclei, metathalamus and epithalamus. The quantitative increase of number was first apparent in the midline structures of thalamus in the first postnatal week. And then TGF-alpha-immunoreactive cells progressively increased throughout diendephalon by postnatal days 15. Adult patterns were reached at postnatal days 20. These results indicate that TGF-alpha-immunoreactive cells were first appeared in thalamic midline structures, increased progressively in the first two postnatal weeks, and followed mediolateral gradient. In addition to maturation of TGF-alpha-immunoreactive cells requires a relatively prolonged period of time to achieve an adult configuration. Also, the early appearance of TGF-alpha immunoreactivity in most diencephalic nuclei may be related to the early appearance of EGFR immunorecativity in many other brain regions. Taken together, these findings suggest that TGF-alpha immunoreactivity correlated with the appearance of the related functional activity in the different regions of diencephalon.

Polyamines Regulate Growth Factor-Induced Protein Phosphorylation in MCF-7 Human Breast Cancer Cells / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Growth factors stimulate protein phosphorylation resulting in transmission of mitogenic signals. In breast cancer, protein kinases and their substrate proteins are importnat in cell proliferation and phathogenesis. Polymine is known as a mediator of stimuli-induced proliferation in many cell systems. In the present study, we report the importance of polyamines in protein phosphorylation in MCF-7 human breast cancer cells. MATERIALS AND METGODS: Protein phosphorylation study was done by incubating cells in the DMEM containing [gamma-(32)P]-ATP. Quantitation of phosphorylation was analysed by fluorescene image analyzer. Tyrosine phosphorylation was detected by anti-phosphotyrosine antibody. Shc was detected by radioimmunoprecipitation and Western blotting. RESULTS: E2, TGF-alpha, and EGF enhanced the protein phosphorylation in very similar pattern. Among those proteins, 67 kDa protein was most strongly phosphorylated. But the most prominent tyrosine phosphoprotein was 52 kDa protein. DFMO at 5 mM strongly inhibited the phosphorylation of the most proteins. Externally added polyamine could recover the inhibitory effect of DFMO in protein phosphorylation. Among the 5 major tyrosine phosphoproteins, 52 and 46 kDa proteins appeared to be Shc proteins. CONCLUSION: Polyamines modulate signal transduction in relation with estrogen receptor and EGF receptor through multiple steps of protein phosphorylations. Tyrosine phosphorylation of Shc proteins were most significantly influenced by polyamines in growth factor-stimulate breast cancer cell proliferation.

Comparison of Transforming Growth Factor-alpha and Epidermal Growth Factor Expression in Melanocytic Nevi: An Immunohistochemical Study / 대한해부학회지

Transforming growth Factor-alpha(TGF-alpha) and epidermal growth factor (EGF) are polypeptides which interact with the epidermal growth factor receptor (EGFR) to produce its biological effects. In normal human tissues, its immunocytochemical presence and biological effects have been demonstrated. The aim of the present investigation was to elucidate the immunolocalization of TGF-alpha and EGF in melanocytic nevi. The data presented in this paper focus attention on comparison of TGF-alpha and EGF in intradermal nevus. The expression of TGF-alpha was stronger than EGF in epidermis of melanocytic nevi. In intradermal nevus, TGF-alpha immunoreactivities were present in most of layers of epidermis and a group of nevus cells in dermis were uniformly immunoreactive with minimal cytoplasm. The expression of EGF was found in part of cytoplasm of keratinocytes in the stratum spinosum and stratum granulosum of epidermis. However, keratinocytes in the stratum basale and nevus cells did not demonstrate immunoreactivity for EGF. In epidermal appendages, the staining intensity of EGF was generally weaker than for TGF-alpha except cells in the external root sheath of hair follicle. In conclusion, some regional variations in the intensity of the immunostaining between TGF-alpha and EGF were present in melanocytic nevi. Our results indicate that TGF-alpha may play a role in growth and proliferation of nevus cells.

Transforming Growth Factor-alpha Regulation of Insulin-like Growth Factor-II, Insulin-like Growth Factor Binding Protein-1 and 3 secretion and Epidermal Growth Factor receptor expression in human luteinized granulosa cells / 대한산부인과학회잡지

OBJECTIVE: To investigate the effects of transforming growth factor (TGF)-alpha on insulin-like growth factor (IGF)-II, insulin-like growth factor binding protein (IGFBP)-1, and 3 secretion and epidermal growth factor (EGF) receptor expression in cultured human luteinized granulosa cells. MATERIALS AND METHODS: Human luteinized granulosa cells were obtained from follicular fluid by transvaginal oocyte aspiration from infertile patients undergoing in vitro fertilization and cultured for 72 hours with TGF-alpha at concentration of 1.0, 10.0, 100.0 ng/ml. The luteinized granulosa cells not treated with TGF-alpha served as control. The secretion of IGF-II, IGFBP-1 and 3 were determined in conditioned media by immunoradiometric assay (IRMA) and reverse transcription-polymerase chain reaction (RT-PCR) was performed for EGF receptor mRNA expression. RESULTS: The cell numbers of 1.0 and 10.0 ng/ml supplement groups were significantly decreased compared to control (p<0.05, p<0.05, respectively), although the cell viabilities were similar in all groups. IGF-II levels were significantly higher in TGF-alpha treatment group at 1.0 and 10.0 ng/ml (p<0.01, p<0.01, respectively), but lower in 100.0 ng/ml (p<0.01). However, the concentrations of IGFBP-1, and 3 per one granulosa cell in each group were no statistically significant differences among the groups. The mRNA concentration of EGF receptor in cultured human luteinized granulosa cells were not significantly different among the groups. CONCLUSION: This study suggests that TGF-alpha regulate intrafollicular bioavailable IGF-II levels, by which TGF-alpha might involved luteinizations. However, TGF-alpha may not directly regulate EGF receptor mRNA expression in cultured human luteinized granulosa cells.

Influence of Transforming Growth Factor-alpha on Expression of Matrix metalloproteinase-2 and Matrix metalloproteinase-9 mRNA in Mouse Embryos / 대한산부인과학회잡지

OBJECTIVE: To investigate the influence of transforming growth factor-alpha (TGF-alpha) on the expression of Matrix metalloproteinase-2 (MMP-2) and Matrix metalloproteinase-9 (MMP-9) mRNA in mouse embryos. MATERIALS AND METHOD: Eight-cell stage mouse embryos were cultured for 48hours with TGF-alpha at concentrations of 1, 10 and 100 ng/ml. Embryos not treated with TGF-alpha served as control. Reverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the expression of MMP-2 and MMP-9 mRNA in developed blastocysts. Following reverse transcription, strategically designed nested primers, optimized for specificity, were used for amplification from the cDNA equivalent of a single embryo. The products were then verified by restriction enzyme digestion and sequence analysis. Results were analyzed with analysis of variance (ANOVA) and statistical significance was defined as p<0.05. RESULTS: The relative quantities (relative volume x intensity) of MMP-2 mRNA expressed in embryos of 10 and 100 ng/m of TGF-alpha treatment groups were significantly increased than in the control and 1 ng/ml of TGF-alpha treatment group (67.2+/-7.5 and 77.4+/-11.6 vs. 38.6+/-4.5 and 43.4+/-6.1, p<0.001). The relative quantities of MMP-9 mRNA of 100 ng/ml TGF-alpha treatment group was significantly increased than in the control and 1 ng/ml TGF-alpha treatment groups (67.6+/-6.5 vs. 36.6+/-14.2 and 40.2+/-11.3, p<0.001, p<0.01, respectively). CONCLUSION: This study suggests that TGF-alpha itself may induce the expression of MMP-2 and 9 mRNA in mouse embryos.

Immunocytochemical Localization of Transforming Growth Factor-alpha in the Forebrain of the Rat / 대한해부학회지

The distribution of Transforming growth factor-alpha (TGF-alpha) was examined in the rat forebrain by immunocytochemistry. TGF-alpha immunoreactivity was observed in the cerebral hemispheres, thalamus and hypothalamus. Neurons in the olfactory and septal area, cerebral cortex, hippocampus, striatum, amygdala, and different nuclei of the thalamus and hypothalamus showed immunoreactivity. The intensity of the immunoreaction was high in the hippocampus, pyramidal cell layers of cerebral cortex, reticular and ventral thalamic nuclei, and paraventricular and supraoptic hypothalamic nuclei. In addition, a few labelled glial cells appeared at random in the forebrain. These results indicate that both neurons and glial cells appear to synthesize TGF-alpha in normal forebrain of the rat. However, TGF-alpha immunoreactivity was more widely distributed in neurons than glial cells. Therefore, although the role of TGF-alpha in the central nervous system remains elusive, the present data support the concept that TGF-alpha may act as a trophic factor in the adult rat forebrain.

Effect of octreotide on transforming growth factor-alpha in human hepatocellular carcinoma cells / 中国癌症杂志

Purpose:To investigate the effects of octreotide on TGF-? autocrine in human hepatocellular carcinoma cells SMMC-7721 and TGF-?-induced cells proliferation. Methods:The expression of TGF-?in SMMC-7721 was determined by radioimmunoassay and reverse-transcriptase polymerase chain reaction(RT-PCR). The expression of epidermal growth factor receptor (EGFR) in the cells was determined by immunohistochemistry method and RT-PCR. The proliferative activity of the cells was evaluated by flow cytometry and colony-forming assay. Results:The content of TGF-?was significantly attenuated by octreotide and the inhibitor rate was 15.0%~26.7%. TGF-?mRNA index was decreased by octreotide.TGF-?increased the expression of EGFR both in mRNA and protein level,while octreotide inhibited the expression induced by TGF-?. Proliferative index (PI) and colony-forming rates were obviously lower in octreotide and TGF-?-treated cells than those in TGF-?-treated cells. Conclusions:There exists a TGF-? autocrine loop in human hepatocellular carcinoma cells SMMC-7721. octreotide could inhibit TGF-? autocrine in the cells, and consequently exerts an inhibitory effect on cell proliferation.

Expression of Epidermal Growth Factor, Transforming Growth Factor-alpha and Epidermal Growth Factor Receptor in Human Cervical Neoplasia / 대한산부인과학회잡지

OBJECTIVE: This study seeks to define the expression of epidermal growth factor(EGF) and transforming growth factor(TGF)-alpha and epidermal growth factor receptor(EGFR), and the relationship to the tumor progression of human cervical epithelial neoplasia. METHODS: To confirm the expression of EGF, TGF-alpha and EGFR immunohistochemically in normal cervix, cervical intraepithelial neoplasms and cervical carcinomas, we used monoclonal antibodies to EGF, TGF-alpha and EGFR. RESULTS: Immunohistochemical stainings using anti-EGF, anti-TGF-alpha and anti-EGFR antibodies showed weak or moderate stainings in all cases. Normal and CIN cases showed predominantly basal and parabasal expression of EGF, TGF-alpha and EGFR, and its expression decreased as the cells became increasingly differentiated toward the surface of the epithelium. In the cervical carcinoma EGF and TGF-alpha expressed weakly to moderately focally, and EGFR expressed intensely in all malignant cells. CONCLUSION: These results suggest that EGF, TGF-alpha and EGFR may involved in cellular proliferation of cervical squamous epithelium and have a significant role in the progression of cervical cancer.