Immediate-early gene expression in EGF-stimulated intestinal epithelial cells.
The mammalian small intestinal epithelium is a continuously renewable population of cells which arise from a proliferative zone of undifferentiated stem cells within the crypts. Epidermal growth factor (EGF) is thought to play a role in the maintenance and proliferative response of the intestinal epithelium. In order to investigate the early changes in intestinal gene expression which occur in response to a mitogenic stimulus, we performed studies in two different cell lines (IEC-6 and HT-29), both of which have characteristics of intestinal crypt cells. Cells were grown in DMEM + 10% FCS at 37 degrees C/5% CO2 and treated with either EGF (10 ng/ml) and/or cycloheximide (CHx) (10 micrograms/ml) for various times. Northern blot analyses were performed on total RNA using 32P-labeled cDNA probes corresponding to various protooncogenes. Our results showed that EGF treatment caused rapid increases in c-fos, c-jun, and junB expression (P < 0.001) in both cell lines. c-fos and c-jun followed similar time courses, peaking at 30 min, whereas junB levels plateaued at 1 hr. The magnitude and time course of protooncogene induction by EGF were similar in the two cell lines. A dose-response experiment indicated peak EGF effects at 10 ng/ml. CHx treatment resulted in greater and more prolonged increases in protooncogene expression when compared to EGF alone, indicating that protein synthesis is not required for protooncogene induction.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Surgery, Beth Israel Hospital, Harvard Medical School, Boston, Massachusetts 02215.
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Hodin
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Meng
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Nguyen
D000818:Animals; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D015870:Gene Expression; D017781:Genes, Immediate-Early; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D011519:Proto-Oncogenes; D013936:Thymidine / Q000378:metabolism
D004815:Epidermal Growth Factor; D003513:Cycloheximide; D013936:Thymidine