Liver Epidermal Growth Factor Research Articles

EGF and risk of hepatocellular carcinoma

Functional polymorphism of the epidermal growth factor (EGF) gene increases risk of hepatocellular carcin oma (HCC) in patients with cirrhosis by increasing serum and liver EGF concentrations (JAMA 2008; 299: 53–60). Previous studies have shown that EGF polymorphisms modulate EGF con centrations and, in mouse models, over-expression of EGF has led to development of HCC. “EGF and the EGFR (EGF receptor) pathway appear to be excellent targets for chemo prevention in patients with cirrh osis”, says the study’s lead author Kenneth Tanabe (Massachusetts General Hospital, Boston, MA, USA). “Measure ment of EGF [concentration] and EGF single nucleotide polymorphism is rel evant in patients with cirrhosis to ass ess their risk for HCC and potentially mod ulate intensity of screening.” The researchers studied mol ecular mech anisms that linked 61*G allele polymorphisms and EGF exp ression in HCC cell lines and human liver tissue. They then studied the assoc iation between EGF polymorphisms and HCC in patients with cirrhosis in two case– control studies (a US study [n=207] and a French validation study [n=121]). EGF secretion was 2·3–times higher in G/G cell lines than in A/A cell lines, the researchers noted. In the US study, compared with the A/A genotype, patients with the A/G genotype had an odds ratio (OR) of 2·4 (95% CI 1–5·4; p=0·05) for development of HCC, and for those with the G/G genotype the OR was 4·0 (1·6–9·6; p=0·002). Number of copies of G was signifi cantly associated with HCC, after adjusting for various factors (hazard ratio for G/G plus A/G vs A/A 3·49 [1·29–9·44; p=0·01]). “The results strongly suggest the importance of inhibiting the EGF/ EGFR axis as a therapeutic strategy for HCC patients,” comments Camillo Porta (IRCCS San Matteo Univ ersity Hospital Foundation, Pavia, Italy). This study hints at the opportunity to select patients more appropriately for EGFR antagonists, adds Ulrich Lehmann (Hannover Medical School, Hannover, Germany).

Epidermal Growth Factor Gene Functional Polymorphism and the Risk of Hepatocellular Carcinoma in Patients With Cirrhosis

Overexpression of epidermal growth factor (EGF) in the liver induces transformation to hepatocellular carcinoma in animal models. Polymorphisms in the EGF gene modulate EGF levels. To assess the relationship among human EGF gene single-nucleotide polymorphism, EGF expression, and risk of hepatocellular carcinoma. Molecular mechanisms linking the 61*G allele polymorphism to EGF expression were examined in human hepatocellular carcinoma cell lines and human liver tissue. A case-control study involving 207 patients with cirrhosis was conducted at the Massachusetts General Hospital (1999-2006) and a validation case-control study involving 121 patients with cirrhosis was conducted at Hôpital Paul Brousse (1993-2006). Restriction fragment-length polymorphism was used to determine the EGF gene polymorphism genotype. Logistic regression analysis was used to assess the association between the EGF polymorphism and hepatocellular carcinoma risk. Mechanisms by which the EGF gene polymorphism modulates EGF levels and associations among EGF gene polymorphism, EGF levels, and hepatocellular carcinoma. Transcripts from the EGF 61*G allele exhibited more than a 2-fold longer half-life than those from the 61*A allele, and EGF secretion was 2.3-fold higher in G/G hepatocellular carcinoma cell lines than A/A cell lines. Serum EGF levels were 1.8-fold higher in G/G patients than A/A patients, and liver EGF levels were 2.4-fold higher in G/G patients than A/A patients. Among the 207 patients with cirrhosis in the Massachusetts study population, 59 also had hepatocellular carcinoma. Analysis of the distribution of allelic frequencies revealed that there was a 4-fold odds of hepatocellular carcinoma in G/G patients compared with A/A patients in the Massachusetts study population (odds ratio, 4.0; 95% confidence interval [CI], 1.6-9.6; P = .002). Logistic regression analysis demonstrated that the number of copies of G was significantly associated with hepatocellular carcinoma after adjusting for age, sex, race, etiology, and severity of cirrhosis (G/G or A/G vs A/A; hazard ratio, 3.49; 95% CI, 1.29-9.44; P = .01). The significant association was validated in the French patients with alcoholic cirrhosis and hepatocellular carcinoma. The EGF gene polymorphism genotype is associated with risk for development of hepatocellular carcinoma in liver cirrhosis through modulation of EGF levels.

Appearance of exogenous epidermal growth factor in liver, bile, and intestinal lumen of suckling rats

This study was performed to investigate the distribution and the degradation of IV administered [125I]rat epidermal growth factor (rEGF) in the liver and gastrointestinal tract of suckling rats. The bile duct of anesthetized rats was cannulated, and [125I]rEGF was injected (with or without 2500-fold excess unlabeled rEGF) into the femoral vein. After 5, 30, 60, and 120 minutes, the radioactivity in the liver, stomach, small intestine, blood, kidney, bile, and luminal contents of the stomach and small intestine was measured. The extracted radioactivity was then analyzed by immunoaffinity chromatography and binding to EGF-specific receptors. High levels of radioactivity were found in the liver (57% of total administered) and small intestine (10%) at 5 minutes, which gradually decreased. On the contrary, radioactivity secreted in the bile and luminal contents of the small intestine increased with time. The radioactivity in the bile represented 2.4% and 4.5% of the total administered at 60 and 120 minutes, respectively. During the first 60 minutes, more than 90% of the radioactivity in the liver, small intestine, bile, and intestinal contents was immunoreactive. Thirty-four to seventy percent of the radioactivity in the bile and liver and 20%–41% of radioactivity in the small intestinal wall and contents were capable of binding to EGF-specific receptors. Radioactivity detected in the liver, bile, small intestine, and intestinal contents was profoundly reduced by the coinjection excess of unlabeled EGF. These studies show that IV administered [125I]rEGF is rapidly taken up by the liver and the gastrointestinal tract and secreted into the bile and intestinal luminal contents of suckling rats in form(s) capable of binding to anti-EGF antibody and EGF-specific receptors. The uptake and secretion by the liver and the small intestine appear to be receptor mediated.

Ontogenesis of epidermal growth factor in liver of BALB mice.

To characterize the ontogenesis of hepatic epidermal growth factor (EGF) metabolism in normal BALB mice, we measured serum and liver concentrations of EGF and liver concentrations of pre-pro EGF mRNA. Female and male animals were studied at 1, 2, 5, 7, and 10 wk of life. After death, body weight and length were measured, and serum and liver tissues were collected for EGF determinations. Immunoreactive serum EGF (means +/- SE) increased at 7 and 10 wk and was significantly higher (P less than 0.05) in males (465 +/- 58 and 683 +/- 120 pg/ml) than females (188 +/- 52 and 295 +/- 64 pg/ml). Liver EGF concentrations were low at 1, 2, and 5 wk, significantly increasing (P less than 0.01) at 10 wk to 179 +/- 36 vs. 268 +/- 49 pg/mg protein for females and males, respectively (female and male values were significantly different, P less than 0.01). Pre-pro EGF mRNA was examined at 1, 2, 3, 5, 7 and 10 wk. EGF message increased in liver to highest values at 10 wk in both males and females. There was a high correlation between serum and liver EGF concentrations during the first 10 wk (r = 0.97 and 0.85 for males and females, respectively) and a twofold increase in liver EGF mRNA between 3 and 10 wk of postnatal life. These results suggest that liver may be an important source of circulating EGF in developing BALB mice.

Decreased levels of hepatic epidermal growth factor receptors in obese hyperglycemic rodents.

Stimulation of epidermal growth factor (EGF) receptor autophosphorylation by EGF and phosphorylation of a Mr 52,000 protein endogenous to the membrane extracts were decreased 6-12-fold in liver membrane extracts from mice homozygous for either the ob/ob or db/db mutation when compared to controls. Liver membranes from the mutant mice bound 4-5-fold less 125I-EGF/unit of protein than did their normal littermates, but exhibited normal EGF binding affinity. Similar decreases in EGF binding were noted in liver membranes from homozygous fa/fa Zucker rats, another obese, hyperinsulinemic animal model, when compared to values from control animals. We also immunoprecipitated hepatic EGF receptors from mice injected with [35S]methionine, and found that livers from db/db mice contained approximately 35% of the labeled EGF receptors found in control animals. Both ob/ob and db/db mice had serum immunoreactive EGF levels similar to or lower than those found in unaffected littermates, suggesting that ligand-mediated down-regulation of receptors was not the cause of the decreased EGF binding. In one mutant, db/db, the decreased binding was associated with a 6-fold decrease in the levels of liver EGF receptor mRNA transcripts; in the ob/ob mice, at most a 2-fold decrease in the level of liver EGF receptor transcripts was observed. EGF binding to cultured peritoneal fibroblasts derived from db/db mice was normal, suggesting that the abnormality in the mutant mice might result from altered environmental or tissue-specific factors rather than an abnormal receptor gene. This was supported by Southern blot analysis of DNA from these animals, which showed identical restriction fragment patterns for the EGF receptor gene in both control and mutant animals. These data indicate that three distinct strains of obese hyperglycemic rodents have decreased levels of hepatic EGF receptors, and suggest that this decrease may result from altered environmental stimuli or tissue-specific factors rather than a primary defect in the EGF receptor gene.

Epidermal growth factor mimics insulin effects in rat hepatocytes.

Epidermal growth factor (EGF) mimicked the effect of insulin to activate glycogen synthase and stimulate glycogen synthesis in isolated rat hepatocytes. Both agents required glucose (greater than 5 mM) and had similar time courses of action. The maximum effect of EGF was approx. 70% of that of insulin, and the half-maximally effective concentrations were 9 nM and 4 nM respectively. Combinations of the two agents produced additive responses. EGF also resembled insulin in its ability to inhibit the effects of 0.1-1.0 nM-glucagon on cyclic AMP and glycogen phosphorylase in hepatocytes. The maximum effect of EGF was approx. 70% of that of insulin, and the half-maximally effective concentrations were approx. 5 nM and 0.5 nM respectively. EGF and insulin inhibited phosphorylase activation by exogenous cyclic AMP, and inhibited cyclic AMP accumulation induced by forskolin. They also inhibited phosphorylase activation provoked by phenylephrine, but not by vasopressin. EGF added alone rapidly activated phosphorylase and increased cytosolic [Ca2+], but the effects were no longer apparent at 5 min and were smaller than those of vasopressin. Insulin did not induce these changes. In hepatocytes previously incubated with myo-[3H]inositol, EGF did not significantly increase myo-inositol 1,4,5-trisphosphate. However, its ability to increase cytosolic [Ca2+] was blocked by neomycin, an inhibitor of phosphatidylinositol bisphosphate hydrolysis. It is concluded that some, but not all, of the effects of EGF in liver are strikingly similar to those exerted by insulin, suggesting that these agents may have some similar mechanisms of action in this tissue.