Newborn rabbit gastric smooth muscle cell culture: EGF and TGF-alpha are potent mitogens.
Newborn rabbit gastric smooth muscle cell culture: EGF and TGF-alpha are potent mitogens.
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新生兔胃平滑肌细胞培养：EGF 和 TGF-α 是有效的有丝分裂原。
DOI：
10.1097/00005176-199308000-00005
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发表时间：
1993
影响因子：
2.9
通讯作者：
Hyman,PE
中科院分区：
文献类型：
--
作者：
Yuan,QX;McRoberts,JA;Lakshmanan,J;Yagi,H;Hyman,PE
表皮生长因子(EGF)和转化生长因子-α(TGF-α)是许多胃肠道上皮生长促进肽家族中的两种。本研究旨在评估它们对培养的胃肌细胞的促有丝分裂作用，并表征这些细胞上的特定 EGF 受体。从新生兔胃底分离单个肌细胞并置于组织培养中。通过用平滑肌肌动蛋白特异性单克隆抗体(CGA7)进行免疫染色评估，汇合时培养物的组成为> 95%肌细胞。为了评估假定的生长因子的作用，将新鲜分离的肌细胞在存在或不存在生长因子的情况下在含有1%胎牛血清的Dulbecco's改良Eagle's (DME)培养基中孵育。 6天后，在持续存在或不存在生长因子的情况下，将细胞在含有[3H]胸苷(1μCi/ml)的无血清培养基中孵育。 24小时后，EGF和TGF-α而非胰岛素样生长因子I (IGF-1)诱导[3H]胸苷掺入呈剂量依赖性增加。十纳摩尔EGF或TGF-α使[ 3 H]胸苷掺入增加超过对照六倍以上。 EGF比TGF-α更有效，表观中位有效剂量(ED50)值分别为64+/-14pM和166+/-62pM(p<0.05)。 EGF 与培养的肌细胞结合，Kd= 7.6+/-1.8 nM，Bmax= 27+/-11 pmol/mg DNA 或 440,000 个受体/细胞。 TGF-α竞争结合这些受体。尽管 IGF-I 不刺激胸苷掺入，但在胃肌细胞上检测到 IGF-I 的特异性高亲和力受体。这些结果表明EGF家族可能在胃平滑肌的生长中发挥作用。
Epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-[alpha]) are two in a family of growth-promoting peptides for many gastrointestinal epithelia. This study was designed to assess their mitogenic effect on cultured gastric myocytes and to characterize specific EGF receptors on these cells. Single myocytes were isolated from newborn rabbit gastric fundus and placed into tissue culture. The composition of the culture at confluence as assessed by immunostaining with smooth muscle actin-specific monoclonal antibody (CGA7) was> 95% myocytes. To assess the effect of putative growth factors, freshly isolated myocytes were incubated in Dulbecco's modified Eagle's (DME) medium containing 1% fetal bovine serum in the presence or absence of growth factors. After 6 days, cells were incubated in serum-free medium with [3H] thymidine (1 [mu] Ci/ml) in the continued presence or absence of growth factors. After 24 h, EGF and TGF-[alpha] but not insulin-like growth factor I (IGF-1) induced a dose-dependent increase in [3H] thymidine incorporation. Ten nanomolar EGF or TGF-[alpha] increased [3H] thymidine incorporation more than sixfold over control. EGF was more potent than was TGF-[alpha], with apparent median effective dose (ED50) values of 64+/-14 pM and 166+/-62 pM (p< 0.05), respectively. EGF bound to cultured myocytes with Kd= 7.6+/-1.8 nM and Bmax= 27+/-11 pmol/mg DNA or 440,000 receptors/cell. TGF-[alpha] competed for binding at these receptors. Although IGF-I did not stimulate thymidine incorporation, specific high-affinity receptors for IGF-I were detected on gastric myocytes. These results suggest that the EGF family may play a role in the growth of gastric smooth muscle.