Overexpression Of Epidermal Growth Factor Research Articles

Synthesis, Characterization and Docking Studies of Novel Series of Benzothiazoles for Anticancer Activity

Background: Cancer is the main cause of mortality worldwide, accounting for the majority of deaths. There is a therapeutic need for innovative, selective anti-tumor medicines that avoid most of the undesirable negative reactions linked to current chemotherapy regimens. The receptor for epidermal growth factor (EGFR) is a membrane glycoprotein, and mutations that cause EGFR overexpression or overactivity have been linked to a range of human malignancies. Objective: In light of this, benzothiazole compounds were developed and produced as EGFR inhibitors, which could possibly form a new class of successful cancer therapies. Experimental work: In this study, we synthesized novel [4-(1H-benzothiazol-2-yl) phenyl] amides derivatives. Furthermore, these produced compounds were subjected to molecular docking experiments to elaborate on interaction and chemistry in order to determine their affinity for the target. After confirming their molecular makeup, these compounds were tested for anticancer efficacy. Result and Discussion: Benzothiazole molecules were effectively synthesized, and their molecular structures were verified using protons NMR and IR spectra. The cytotoxic activity of these novel compounds was determined using the SRB technique. Conclusion: The present investigation involves the targeted chemotherapy to plausibly reverse, suppress or prevent cancer. Synthesized compounds revealed good cytotoxic effect and have shown more potential. So, in future, these effective molecules will be explored for increasing their affinity towards different targets, so they can be used against various cancers.

Paget’s disease of the breast: a contemporary perspective

The treatment strategy for Paget’s breast cancer (PBC), as for other morphological forms, is determined by the spread of the invasive tumor process, as well as the absence or presence of adverse factors. Significant prognostic factors, e.g. the invasive component of Paget’s cancer, the involvement status of regional lymph nodes, morphological criteria, the grade of malignancy, overexpression of epidermal growth factor Her2/neu, the presence of BRCA 1/2 and CHEK2 gene mutations, as well as age, affect the prognosis of Paget’s disease. According to various sources, PBC in 90–98 % is combined with invasive or non‑invasive breast cancer. For the most part, the lesion is multifocal. There’s high expectation for PBC to form tumor node. A palpable tumor node in the mammary gland is detected in half of the patients. PBC has certain features and specificity in relation to treatment methods. The method of radical surgical treatment of patients is primarily based on performing oncoplastic resections. Sufficient breast volume is a significant criterion for the possibility of performing an organ‑ preserving operation. The organ‑ preserving treatment is oncologically safe in case of PBC. The removal of the nipple‑ areolar complex is mandatory in case of PBC. It is possible to improve survival statistics by analyzing the biological characteristics of the tumor and developing a more accurate approach to the treatment of patients in this category. Determining clear indications for organ‑p reserving operations is the main opportunity to improve the quality of life of patients and their further rehabilitation.

Autocrine EGF and TGF-α promote primary and acquired resistance to ALK/c-Met kinase inhibitors in non-small-cell lung cancer.

Drug resistance severely limits the clinical therapeutic value of molecularly targeted drugs. Growth factors gain a tremendous amount of focus due to the ability to promote drug resistance in non-small-cell lung cancer (NSCLC). However, whether tumor cells themselves can mediate drug resistance by secreting growth factors needs further clarification. Here, we first screened growth factors to identify autocrine epidermal growth factor (EGF) and transforming growth factor alpha (TGF-α) that caused primary resistance to the ALK inhibitor TAE684 in H3122 cells and the c-MET-specific inhibitor SGX-523 in EBC-1 cells. Next, we discovered increased autocrine production of EGF and TGF-α in established acquired resistant H3122/TR and EBC-1/SR cells. Importantly, overexpression of EGF and TGF-α in two NSCLC cell lines produced resistance to TAE684 and SGX-523. Clinically, NSCLC patients with high expression of EGF and TGF-α developed primary resistance to crizotinib. Mechanistically, autocrine EGF and TGF-α activated EGFR signaling pathways to survive targeted c-Met and ALK inhibition. Furthermore, combined treatment with gefitinib circumvented EGF- and TGF-α-mediated primary and acquired resistance to TAE684/SGX-523. Taken together, these results suggested increased autocrine EGF and TGF-α conferred primary and acquired resistance to ALK/c-Met kinase inhibitors in NSCLC.

Molecular-genetic mechanisms of the signal cascade RAS-RAF-MEK-ERK associated with the development of the tumor process and the purpose of targeted drugs for colorectal cancer

Colorectal cancer occupies one of the leading positions in the world in the structure of cancer incidence. The vital processes of cancer cells largely depend on the production of growth factors and their receptors. One of these is epidermal growth factor (EGFR), which is a tyrosine kinase receptor for cell membranes. Normally, binding of EGFR ligands and transforming growth factor alpha (TGFα) induces receptor activation, which triggers ERK and PI3K signaling pathways that control cell proliferation, migration, invasion, and many other processes. It was found that in 80% of cases, colorectal cancer occurs as a result of EGFR overexpression, which leads to increased growth and division of tumor cells due to hyperactivation of the RAS-RAF-MEKERK signaling cascade. The RAS-RAF-MEK-ERK cascade is a pathway that regulates cell proliferation, cell cycle, and cell migration. In the development of human cancer, mutations of the RAS/RAF family are most often the cause of dysregulation of signal transduction through this pathway. According to current data, about a third of all malignant neoplasms are associated with mutations in the genes of the RAS family, which include HRAS, KRAS, NRAS, RRAS, and other homologous proteins. Proteins of the RAS family are involved in the activation of tyrosine kinase signaling pathways, which leads to gene mutations. This process determines proliferative activity, ability to differentiate, metastasis, avoidance of apoptosis, induction of angiogenesis. Permanent RAS activation leads to malignant cell degeneration. Thus, the expression and mutation of the EGFR gene are associated with various variants of tumor progression and poor prognosis in malignant neoplasms of various localizations. Over the past decades, significant advances have been made in the treatment of metastatic colorectal cancer. However, the expansion of the spectrum of effective anticancer drugs also creates a number of difficulties in choosing the optimal drug therapy regimens in patients with metastases of colorectal cancer.

A five-year follow up study of stage I-IV of rectal cancer with an emphasis on epidermal growth factor over-expression.

The key pro-proliferative pathway, based on EGFR-KRAS/BRAF-myc, is seen as the main goal of personalized therapy in rectal cancer. The objective of the study is to assess the EGFR immunoreactivity in rectal cancer and to estimate its relationship with the clinical outcome, especially as a predictor of poor outcomes. Patients: applying exclusion criteria, 102 patients with stage I-IV rectal cancer, who had undergone scheduled surgery during the period 2005-2011, were included in the study. There was a follow-up study with a span of 5 years from the date of the surgery. Immunohistochemistry using EGFR (EGFR Ab10, Clone111.6) was performed to detect an overexpression of the targeted receptor. Digital analysis of positive reactions of membranes was performed utilizing VisiopharmTM. The degree of EGFR intensity (log OR 0.854, OR 2.35, 95% Cl: 1.14-4.85, p = 0.021) is a significant factor in the prognosis of death within 2 years of surgery. The OS curve showed a significant decrease after 40 months from the date of surgery in the cases where EGFR had a high expression. The ROC curve for the cancer stage, according to the UICC classification and EGFR expression, in order to predict a 2-year RFS, reached a high specificity value (ROC = 0.81, p = 0.0408). Immunohistochemical EGFR expression is inexpensive, specific and broadly available.

A quiescent state following mild sensory arousal in Caenorhabditis elegans is potentiated by stress

An animal’s behavioral and physiological response to stressors includes changes to its responses to stimuli. How such changes occur is not well understood. Here we describe a Caenorhabditis elegans quiescent behavior, post-response quiescence (PRQ), which is modulated by the C. elegans response to cellular stressors. Following an aversive mechanical or blue light stimulus, worms respond first by briefly moving, and then become more quiescent for a period lasting tens of seconds. PRQ occurs at low frequency in unstressed animals, but is more frequent in animals that have experienced cellular stress due to ultraviolet light exposure as well as in animals following overexpression of epidermal growth factor (EGF). PRQ requires the function of the carboxypeptidase EGL-21 and the calcium-activated protein for secretion (CAPS) UNC-31, suggesting it has a neuropeptidergic mechanism. Although PRQ requires the sleep-promoting neurons RIS and ALA, it is not accompanied by decreased arousability, and does not appear to be homeostatically regulated, suggesting that it is not a sleep state. PRQ represents a simple, tractable model for studying how neuromodulatory states like stress alter behavioral responses to stimuli.

The Overexpression of Epidermal Growth Factor (EGF) in HaCaT Cells Promotes the Proliferation, Migration, Invasion and Transdifferentiation to Epidermal Stem Cell Immunophenotyping of Adipose-Derived Stem Cells (ADSCs)

Background and ObjectivesThe application of adipose derived stem cells (ADSCs) in skin repair has attracted much attention nowadays. Epidermal growth factor (EGF) participates in the progress of skin proliferation, differentiation and so forth. We aimed to explore the role of EGF in the proliferation, invasion, migration and transdifferentiation into epidermal cell phenotypes of ADSCs.Methods and ResultsADSCs were extracted from adipose tissues from patient. Immunophenotyping was determined by flow cytometry. Overexpressed EGF or siEGF was transfected by lentiviruses. EGF was determined by enzyme linked immunosorbent assay (ELISA) or western blot. ADSCs and HaCaT cells were co-cultured by Transwell chambers. Conditioned medium (CM) was obtained from cultured HaCaT cells and used for the culturing of ADSCs. Cell viability was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Invasion rate was measured by Transwell invasion assay and migration rate by wound healing test. mRNA and protein levels were measured by qPCR and western blot respectively. The extracted cells from adipose tissues were identified as ADSCs by morphology and immunophenotyping. The expression of EGF was up or down regulated constantly in HaCaT cell line after transfection. EGF overexpression upregulated the proliferation, migration and invasion rates of ADSCs, and EGF expression regulated the expression of cytokeratin-19 (CK19) and integrin-β as well.ConclusionsEGF could be served as a stimulus to promote the proliferation, migration, and invasion as well as the transdifferentiation into epidermal stem cell immunophenotyping of ADSCs. The results showed that EGF had a promising effect on the repair of skin wound.

DUSP2 methylation is a candidate biomarker of outcome in head and neck cancer.

Biomarkers predictive of response to chemoradiotherapy (CRT) regimens for locally advanced head and neck squamous cell carcinoma (LA-HNSCC) are urgently required to identify patients in whom this approach is likely to be effective. TP53 mutations and epidermal growth factor (EGFR) overexpression are common markers of disease. Dual-specificity-phosphatase-2 (DUSP2) has an essential role in cell proliferation, cancer and immune responses. Aberrant DUSP2 methylation was investigated by pyrosequencing in 5 HNSCC cell lines, 112 LA-HNSCC tumours. EGFR was investigated by immunohistochemistry and TP53 was analysed by sequencing. We demonstrate methylation-dependent transcriptional silencing of DUSP2 in HNSCC cell lines. In LA-HNSCC patients, aberrant methylation in the DUSP2 CpG island was present in 51/112 cases (45.5%). LA-HNSCC cases with wild-type TP53, overexpression of EGFR and unmethylated DUSP2 had the worst overall survival (P≤0.001). DUSP2 methylation, when combined with EGFR and TP53, is a candidate biomarker of clinical outcome in LA-HNSCC treated with CRT.

EGF Level in Hepatoid Gland Adenomas and Hepatoid Gland Epitheliomas in Dogs After Administering Tamoxifen.

Neoplastic lesions of perianal glands account for approximately 10% of all skin cancer cases in dogs. They occur in many dog breeds, usually in male animals aged over 6 years. Due to their hormone-dependency, tamoxifen can be used in antineoplastic treatment. The aim of the study was to measure epidermal growth factor (EGF) levels in the serum of dogs with perianal tumours after tamoxifen treatment and to use it as a prognostic factor for further treatment. The study was performed on 19 male dogs aged between 6 and 14 years, diagnosed with neoplastic hyperplasia in the perianal region. The control group comprised 10 healthy dogs brought in for routine castration. The research material comprised blood drawn from the animals and tumour specimens for histopathology. The study group received 1-month treatment with tamoxifen. Blood serum was then tested for 17-β oestradiol level, and for EGF level on the first day of the therapy and 6 months after treatment completion. Hepatoid gland adenomas were diagnosed in 10 cases, and hepatoid gland epitheliomas in nine cases. Elevated 17-β oestradiol levels were observed in all dogs. On the first day of treatment with tamoxifen, the serum EGF levels in all study groups were higher than in the control group. At the 6-month follow-up, the EGF levels were significantly reduced in hepatoid gland adenoma cases compared to those taken on the first day of treatment of tamoxifen, while in animals with hepatoid gland epithelioma, it was greatly increased and was correlated with relapse. Perianal gland tumours are characterised by EGF overexpression, which can be helpful in early-stage prognosis and treatment. An increase in EGF levels 6 months after tamoxifen therapy correlates with disease progression and may be a useful prognostic factor.

PWE-039 The Interaction between The Hypoxia Inducible Factor-1Α (HIF-1Α) and ERBB Signalling Pathways in Colorectal Liver Metastases (CRLM)

Introduction Tumour hypoxia is associated with reduced survival by activating multiple molecular pathways such as, but not exclusively, HIF-1α and mTOR/AKT/PI3K pathways. It is also known to cause overexpression of Epidermal Growth Factor (EGFR)/HER1 in human cancer. HER2 (neu) signalling increases the rate of Hypoxia-Inducible Factor 1α (HIF-1α) synthesis. Overexpression of Erbb3/HER3 and Erbb4/HER4 is associated with a higher recurrence rate of stage 2 and 3 colorectal cancer. The aim of this study was to analyse the relationship between the expression of HIF-1α and members of the ErbB pathway in CRLM utilising a novel proteomic based collaborative Enzyme Enhanced Reactive (CEER) immunoassay. Methods Protein lysates of from fresh frozen biopsies and formalin fixed paraffin sections of CRLM from 18 patients who had not undergone neoadjuvant chemotherapy prior to a potentially curative hepatectomy for CRLM were analysed. When multiple metastases were present biopsies and sections were obtained from the largest metastasis. CEER immunoassay was performed on the protein lysates to quantify the phosphorylated and non-phosphorylated forms of receptors; ErbB family receptors (HER1/EGFR, HER2/c-neu, HER3, HER4) and cMET, and phosphorylated downstream ErbB pathway factors AKT, PRAS, RSP6, ERK, and RSK, and CK. Immunostaining was performed on formalin-fixed-paraffin-embedded CRLM sections for HIF-1α. ErbB pathway factor quantification was segregated according to the HIF-1α expression status; HIF-1α +ve and HIF-1α -ve Results HIF-1α was expressed in 15 (83%) of tumours. There was no significant difference between the expression of phosphorylated ‘receptors’, and phosphorylated forms AKT, PRAS, RSP6, ERK, MEK, and PI3K between the HIF-1α +ve and HIF-1α –ve groups. HER3 and IGF1R were higher; (median µm: range; 87 [74–344] vs 222 [60–404]) and (115 [85–206] vs 186 [90–285]) respectively in the HIF-1α +ve group, HER1, HER2, PI3K and CK levels were lower; (108 [40–316] vs 36 [22–46]), (698 [523–870] vs 481 [288–709]), (5338 [3334–6689] vs 2719 [1790–4193]), (5383 [1826–9799] vs 2719 [1789–4193]) respectively in the HIF-1α +ve group. Although a trend was observed, the difference did not reach statistical significance. Conclusion HIF-1α may have a role in the overexpression of HER3 and IGF1R overexpression in CRLM. More research is needed to establish this interrelationship which would help in molecular based therapeutics for CRLM. Disclosure of Interest None Declared

The effect of infrared radiation on the expression of epidermal growth factor and its receptor in atrophic gastritis

Objective To study the effect of infrared irradiation on the expression of epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) in the gastric mucosa of rats with chronic atrophic gastritis (CAG), and to explore its mechanism. Methods Forty healthy adult and clean Wistar male rats were randomized into a normal control group (N group), a model control group (M group) and an infrared group (I group) using a random number table. CAG was induced in the rats of the M and I groups by intra-gastric infusion of 2 ml of a mixture of 30% alcohol and 2% sodium salicylate once every day for 8 weeks. This was combined with compulsory running and irregular fasting as pathogenic factors. After the CAG model had been established, the rats in the I group were given infrared irradiation for 10 min once every day for 20 days. Then all of the rats were sacrificed and the expression of EGF and EGFR was observed using immunohistochemistry. Results The positive staining scores for EGF and EGFR of the M group were 4.91±2.58 and 6.75±2.63, significantly higher than those of the N group (2.10±0.65 and 4.20±2.24 respectively). Those of the I group were 2.75±1.01 and 4.75±1.66, significantly lower than those of the M group, though not significantly different from the N group's averages. Conclusion Infrared irradiation can inhibit over-expression of EGF and EGFR in the gastric mucosa of rats with chronic atrophic gastritis. This should protect the gastric mucosa and promote tissue repair and healing. Key words: Infrared irradiation; Chronic atrophic gastritis; Epidermal growth factor; Epidermal growth factor receptor

Abstract LB-268: Black rapsberries induced protection in pancreatic cancer mouse models

Abstract Backgrounds and objectives Pancreatic ductal adenocarcinoma (PDAC) is the 12th most common cancer worldwide with 338,000 new cases diagnosed in 2012. The estimated 5-year survival rate of diagnosed PDAC patients is less than 4%, making it the 7th leading cause of cancer death. Over-expression of epidermal growth factor (EGF) receptors and ErbB2 receptors has been associated with the development and progression of human PDAC. Diets rich in fruits and vegetables have been reported to reduce the risk of PDAC incidence and its recurrence. Our current study aimed to investigate the effects of dietary black raspberries (BRBs) on pancreatic carcinogenesis.. Methods 4-5 weeks old pancreas-specific (Pdx-1-cre) mutant mice that carry constitutively-activated oncogene K-ras (G12D) and loss-of-function tumor-suppressor gene Trp53 (R172H), hereafter refers to KPC mice, were fed with control (n = 26) or 5% BRB (n = 29) diet. In addition, orthotopic pancreatic cancer xenografts were injected with luciferase transfected human Panc-1 cells. Four weeks after Panc-1 cells were injected, these mice were given control (n = 8) or 5% BRB (n = 19) diet for 6 weeks. Results Kaplan-Meier survival analysis showed that BRBs prolonged survival of KPC mice with a medium survival of 124 days in comparison to 79 days from mice fed with control diet (p = 0.03). Molecular studies suggest that BRBs decreased the activation of EGF receptor and ErbB2 receptor signaling pathways. Further, BRBs significantly suppressed growth of Panc-1 tumors and it was associated with decreased EGF receptor activation. Conclusion These results suggest that BRBs might protect against carcinogenesis of PDAC through regulating EGF receptor and ErbB2 receptor signaling pathways. Further investigation of the downstream pathways of EGF receptor and ErbB2 receptor activation is warranted. Citation Format: Pan Pan, Chad Skaer, Hsin Tzu Wang, Susan Tsai, Kiyoko Oshima, Yi-Wen Huang, Gary Stoner, John Lechner, Li-Shu Wang. Black rapsberries induced protection in pancreatic cancer mouse models. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr LB-268. doi:10.1158/1538-7445.AM2015-LB-268

An essential role of cAMP response element-binding protein in epidermal growth factor-mediated induction of sodium/glucose cotransporter 1 gene expression and intestinal glucose uptake

The sodium/glucose cotransporter 1 (SGLT1) is responsible for glucose uptake in intestinal epithelial cells. Its expression is decreased in individuals with intestinal inflammatory disorders and is correlated with the pathogenesis of disease. The aim of this study was to understand the regulatory mechanism of the SGLT1 gene. Using the trinitrobenzene sulfonic acid-induced mouse models of intestinal inflammation, we observed decreased SGLT1 expression in the inflamed intestine was positively correlated with the mucosal level of epidermal growth factor (EGF) and activated CREB. Overexpression of EGF demonstrated that the effect of EGF on intestinal glucose uptake was primarily due to the increased level of SGLT1. We identified an essential cAMP binding element (CRE) confers EGF inducibility in the human SGLT1 gene promoter. ChIP assay further demonstrated the increased binding of CREB and CBP to the SGLT1 gene promoter in EGF-treated cells. In addition, the EGFR- and PI3K-dependent CREB phosphorylations are involved in the EGF-mediated SGLT1 expression. This is the first report to demonstrate that CREB is involved in EGF-mediated transcription regulation of SGLT1 gene in the normal and inflamed intestine, which can provide potential therapeutic applications for intestinal inflammatory disorders.

Abstract 3592: Macrophages confer resistance to cisplatin in MTLn3 and MDA-MB 231 breast cancer cells

Abstract Drug resistance presents a clinical challenge to the treatment of breast cancer. Stromal cells such as macrophages may play a role in drug resistance. The purpose of this study is to examine the interaction between breast cancer cells and macrophages and its effects on drug therapy resistance. The five-year survival for localized hormone sensitive disease is 98% and as low as 15% for triple negative (ER/PR/Her2) and metastatic disease. Most targeted anticancer therapies, as well as cytotoxic therapies, are burdened by the development of cancer cell resistance. It is hypothesized that the tumor microenvironment may confer innate resistance to drug therapy. It is well established that chemotactic factors found in the tumor microenvironment can promote motility and invasion. We hypothesize that tumor associated macrophages can confer drug resistance to breast cancer cells via overexpression of epidermal growth factor (EGF). EGF is overexpressed in many tumor types and is clinically correlated with a poor overall prognosis. Previous studies have shown that EGF is produced by non-tumor cells in the microenvironment; in particular tumor-associated macrophages (TAM) and endothelial cells. We have previously shown that macrophages and tumor cells, via a paracrine loop, are necessary and sufficient for comigration and invasion. Breast cancer cells produce colony stimulating factor 1 (CSF-1) which positively feeds back to increase macrophage expression of EGF. Increased EGF expression in turn leads to tumor growth and invasion. We selected two triple negative breast cancer cell lines, MDA-MB 231 (human mammary adenocarcinoma) and MTLn3 (rat mammary adenocarcinoma) that were transfected to express GFP. Co-culture cell survival assays were performed with macrophages, fibroblasts, and endothelial cells. All tissue culture was completed in DMEM (HyClone) plus 10% FBS. Cells were grown for 24 hours in co-culture, treated with chemotherapy (doxorubicin or cisplatin at a range of concentrations) for 24 hours, and then analyzed at 72 hours using the DXP-10 Calibur analyzer. Prior to analysis, cells were again replated to measure “replating efficiency” i.e. chemoresistant cell survival. We found that coculture with macrophages increased tumor cell survival upon exposure to cisplatin. We conclude that macrophages are an important component of the tumor microenvironment that may play a role in chemotherapy resistance. Citation Format: Joshua K. Sabari, Ramon Cabrera, Jeffrey Segall. Macrophages confer resistance to cisplatin in MTLn3 and MDA-MB 231 breast cancer cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3592. doi:10.1158/1538-7445.AM2014-3592

Esophageal Cancer and the Importance of Epidermal Growth Factor (EGFR)

Esophageal squamous cell carcinoma (ESCC) is one of the most frequent malignancies, worldwide. It is important to find out what prognostic factors can facilitate diagnosis, optimize therapeutic decisions, and improve the survival of these patients. Despite improvements in surgical techniques combined with chemotherapy and/or radiotherapy, the novel therapies such as small molecule inhibitors of tyrosine kinases (TKIs) and humanized monoclonal antibodies (mAbs) are very much needed. On the other hand, neoadjuvant chemotherapy which may improve the outcome is accompanied by toxicity by destruction of normal cells. Side effects may be avoided by developing therapies that specifically target molecular characteristics of tumors. Epidermal growth factor receptor (EGFR) is one of tyrosine kinases receptors widely distributed in human epithelial cell membrane. Genetic polymorphisms in EGFR genes influence cell cycle progression, angiogenesis, apoptosis and metastasis. EGFR mutations are mostly observed in lung tumors; curiously they are more prevalent in Asian women diagnosed with adenocarcinoma. Also, esophageal SCC shows a relatively high incidence of EGFR (33%) and/or HER2 (31%) overexpression. Patients who carry these mutations in EGFR have been founded tending to have a better response to gefitinib, an EGFR-TKI, whereas patients with the wild-type genotype show a better response to conventional chemotherapy. Therefore, finding clinical characteristics and environmental interactions with EGFR can affect on investigations about novel anti-cancer therapies like monoclonal antibodies and gene therapy and studies which identify patients who may benefit from EGFR targeted therapies. Hence, it may be effective on the improvement of prognosis in these patients.

Epidermal growth factor gene polymorphism 61A/G in patients with chronic liver disease for early detection of hepatocellular carcinoma

Overexpression of epidermal growth factor (EGF) in the liver induces transformation into hepatocellular carcinoma (HCC) in animal models. Polymorphisms in the EGF gene modulate EGF levels. To evaluate the effect of EGF gene single nucleotide polymorphism and to assess its correlation with the risk of HCC in patients with chronic liver diseases. The present study included 80 participants divided into four groups: group 1 included 20 asymptomatic healthy control volunteers, group 2 included 20 patients with chronic hepatitis C viral (HCV) infection, group 3 included 20 patients with liver cirrhosis, and group 4 included 20 patients with HCC. For all participants, the following investigations were performed: routine laboratory investigations including complete blood count, liver function tests, sero markers of hepatitis viruses HBsAg, HCV-RNA by quantitative polymerase chain reaction, and α-fetoprotein. DNA was extracted from whole blood for detection of single nucleotide polymorphism of the EGF by polymerase chain reaction, followed by restriction fragment length polymorphism. We found a significant difference between both patients with HCC and HCV versus controls in terms of the G carrier (GG and GA; 80 vs. 40%, P<0.05). In addition, the cirrhotic and chronic hepatitis C patients with GG had three-fold and 2.3-fold odds ratio for developing HCC, respectively. The EGF 61GG genotype might be associated with a high risk for the development of HCC in Egyptian patients with chronic liver disease.

Epidermal Growth Factor Improves SurvivaL and Prevents Intestinal Injury in a Murine Model of Pseudomonas aeruginosa Pneumonia

Mortality from pneumonia is mediated, in part, through extrapulmonary causes. Epidermal growth factor (EGF) has broad cytoprotective effects, including potent restorative properties in the injured intestine. The purpose of this study was to determine the efficacy of EGF treatment following Pseudomonas aeruginosa pneumonia. FVB/N mice underwent intratracheal injection of either P. aeruginosa or saline and were then randomized to receive either systemic EGF or vehicle beginning immediately or 24 h after the onset of pneumonia. Systemic EGF decreased 7-day mortality from 65% to 10% when initiated immediately after the onset of pneumonia and to 27% when initiated 24 h after the onset of pneumonia. Even though injury in pneumonia is initiated in the lungs, the survival advantage conferred by EGF was not associated with improvements in pulmonary pathology. In contrast, EGF prevented intestinal injury by reversing pneumonia-induced increases in intestinal epithelial apoptosis and decreases in intestinal proliferation and villus length. Systemic cytokines and kidney and liver function were unaffected by EGF therapy, although EGF decreased pneumonia-induced splenocyte apoptosis. To determine whether the intestine was sufficient to account for extrapulmonary effects induced by EGF, a separate set of experiments was done using transgenic mice with enterocyte-specific overexpression of EGF (IFABP-EGF [intestinal fatty acid-binding protein linked to mouse EGF] mice), which were compared with wild-type mice subjected to pneumonia. IFABP-EGF mice had improved survival compared with wild-type mice following pneumonia (50% vs. 28%, respectively, P < 0.05) and were protected from pneumonia-induced intestinal injury. Thus, EGF may be a potential adjunctive therapy for pneumonia, mediated in part by its effects on the intestine.

Investigation of convergence between Epidermal Growth Factor and hypoxia on the pro-angiogenic switch in colorectal cancer

Background: Over-expression of Epidermal Growth Factor (EGF) and hypoxia are features of colorectal cancer (CRC), thus the development of novel treatments targeting EGF and its receptors, such as cetuximab. Recent studies uncovered a pro-angiogenic role for EGF but involvement by Vascular Endothelial Growth Factor (VEGF) is unknown. Moreover, it is unknown whether EGF can synergise with hypoxia in CRC. Aim of the study was to investigate the pro-angiogenic effect induced by EGF in combination with hypoxia in CRC.

Enterocyte-specific epidermal growth factor prevents barrier dysfunction and improves mortality in murine peritonitis

Systemic administration of epidermal growth factor (EGF) decreases mortality in a murine model of septic peritonitis. Although EGF can have direct healing effects on the intestinal mucosa, it is unknown whether the benefits of systemic EGF in peritonitis are mediated through the intestine. Here, we demonstrate that enterocyte-specific overexpression of EGF is sufficient to prevent intestinal barrier dysfunction and improve survival in peritonitis. Transgenic FVB/N mice that overexpress EGF exclusively in enterocytes (IFABP-EGF) and wild-type (WT) mice were subjected to either sham laparotomy or cecal ligation and puncture (CLP). Intestinal permeability, expression of the tight junction proteins claudins-1, -2, -3, -4, -5, -7, and -8, occludin, and zonula occludens-1; villus length; intestinal epithelial proliferation; and epithelial apoptosis were evaluated. A separate cohort of mice was followed for survival. Peritonitis induced a threefold increase in intestinal permeability in WT mice. This was associated with increased claudin-2 expression and a change in subcellular localization. Permeability decreased to basal levels in IFABP-EGF septic mice, and claudin-2 expression and localization were similar to those of sham animals. Claudin-4 expression was decreased following CLP but was not different between WT septic mice and IFABP-EGF septic mice. Peritonitis-induced decreases in villus length and proliferation and increases in apoptosis seen in WT septic mice did not occur in IFABP-EGF septic mice. IFABP-EGF mice had improved 7-day mortality compared with WT septic mice (6% vs. 64%). Since enterocyte-specific overexpression of EGF is sufficient to prevent peritonitis-induced intestinal barrier dysfunction and confers a survival advantage, the protective effects of systemic EGF in septic peritonitis appear to be mediated in an intestine-specific fashion.

Functions and therapeutic value of focal adhesion kinase signaling during hepatocellular carcinoma development and progression

We appreciate the article by Wu et al. in a recent issue of HEPATOLOGY.1 In this study, the authors demonstrated that the overexpression of epidermal growth factor–like domain 7 (Egfl7) was closely associated with poor prognosis in hepatocellular carcinomas (HCCs). In addition, they investigated the role of Egfl7 in the development and progression of HCC by silencing its expression via transfecting a specific small interfering RNA in HCC cell lines. Silencing of Egfl7 expression caused no changes in cell growth, even if it resulted in a relevant inhibition of cell migration, which appeared mediated by the phosphorylation of focal adhesion kinase (FAK). All these effects were reverted by the use of an epidermal growth factor receptor (EGFR) inhibitor. Interestingly, the existence of a critical role of Egfl7/FAK pathway in controlling HCC was also demonstrated in a mouse model of intrahepatic and pulmonary metastases. FAK phosphorylation is a critical event in processes of cell migration, adhesion, and growth of several cancer cells.2 The role of FAK in the invasion, metastasis, and prognosis of HCC was not completely unknown. In fact, a previous work reported that the increased messenger RNA expression of FAK was well correlated with tumor size and serum levels of alpha-fetoprotein, indicating an important prognostic value to evaluate the survival of patients with HCC.3 In addition, more recently, FAK and Src have been demonstrated to be overexpressed and activated in HCC tissues.4 However, the mechanism by which FAK may contribute to HCC pathogenesis and progression has still not been elucidated.5 FAK is a tyrosine kinase that upon integrin ligation cross-interacts with Src, enhancing the phosphorylation of downstream targets involved in migration pathways, such as paxillin.6 This mechanism also seems to be conserved in human hepatoma cell lines, but it is still unknown which upstream signaling molecules might be involved in the Src/FAK/paxillin interaction.7 The mechanism proposed by Wu and colleagues is very intriguing. The authors suggest that the regulation of FAK phosphorylation and activity might be influenced by the binding of epidermal growth factor (EGF) to its membrane receptor (EGFR). This hypothesis is not only suggestive of a possible role of EGFR-FAK axis in HCC progression, but also in tumor development. In fact, the EGF-EGFR combination is engaged in extensive cross-talk with other signaling pathways which control cell proliferation and inflammatory response.8 These findings, once again, encourage the use of EGFR inhibitors as potential therapeutic agents during HCC and suggest that FAK might be a possible novel potential target in therapy.9, 10 Finally, we believe that these last considerations should prompt in vivo and in vitro studies to explore anticancer properties of small molecules (e.g., PF-573,228; PF-562,271; and NVP-226) able to antagonize FAK activity in HCC. Anna Alisi Ph.D.*, Clara Balsano M.D. , * Liver Unit, Bambino Gesù Children's Hospital and, Research Institute, Rome, Italy, Department of Internal Medicine, University of L'Aquila, L'Aquila, Italy.