Regulation of protein breakdown by epidermal growth factor in A431 cells.
Addition of epidermal growth factor (EGF) to cultures of A431 human epidermoid carcinoma cells produces an increase in the rate of intracellular protein breakdown that cannot be accounted for by increased proteolysis in lysates from EGF-treated cells. In support of this observation, inhibition of protein synthesis with cycloheximide does not reduce the EGF response in cell monolayers. On the other hand, inhibitors of lysosomal proteolytic function such as leupeptin, vinblastine and especially the weak base, ammonia, are able to block the ability of EGF to increase protein breakdown. Additional results suggest that the EGF effect is mediated via a stimulation of autophagy. First, the autophagocytosis inhibitor, 3-methyladenine, reduces the EGF response, and second, the ability of insulin to inhibit protein breakdown by preventing the formation of autophagic vacuoles is overcome by EGF. Moreover, the actions of inhibitors and competing hormones are similar to those reported for glucagon, a hormone known to increase autophagy. The EGF response on protein breakdown persists for at least 6 h after thorough washing of the A431 monolayers. This result contrasts with the rapid reversal of EGF effects in other cell lines. Examination of the fate of bound EGF in cells washed and incubated for 2 h at 37 degrees C shows that some 500-fold more EGF per mg protein is retained on the surface of A431 cells compared to AG2804-transformed fibroblasts, a difference which probably explains the unusual persistence of the EGF effect on protein breakdown.
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000643:Ammonium Chloride / Q000494:pharmacology; D001329:Autolysis; D002294:Carcinoma, Squamous Cell; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000378:metabolism / Q000494:pharmacology*; D005347:Fibroblasts; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007328:Insulin / Q000494:pharmacology; D007976:Leupeptins / Q000494:pharmacology; D008168:Lung; D011506:Proteins / Q000378:metabolism*; D013997:Time Factors; D014747:Vinblastine / Q000494:pharmacology
D007328:Insulin; D007976:Leupeptins; D011506:Proteins; D000643:Ammonium Chloride; C025946:3-methyladenine; D014747:Vinblastine; D004815:Epidermal Growth Factor; D003513:Cycloheximide; C032854:leupeptin; D000225:Adenine