Biochemical and Biophysical Research Communications
Amphiregulin is a potent mitogen for the vascular smooth muscle cell line, A7r5
Section snippets
Methods
Cell line and cell culture. A7r5, immortal embryonic rat thoracic aorta smooth muscle cell line, was obtained from the American Type Culture Collection (ATCC No. CRL1444). The A7r5 cell line was grown in Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal calf serum (FCS) under 5% CO2 in air at 37 °C in a humidified incubator.
Reagents. p42/p44-MAPK, p38-MAPK, JNK, phospho-p42/p44-MAPK, phospho-p38-MAPK antibodies, and phospho and nonphospho-Akt/PKB antibodies were from Cell Signaling
Expression of the EGF family and the EGF receptor family in A7r5 cells
It is generally believed that the EGF family plays an important role in cell proliferation, survival, migration, and differentiation. These factors exert their cellular effects through binding to cell surface receptors. The EGF family of receptors include epidermal growth factor receptor (EGFR also called ErbB1/HER1) [15], ErbB2 (Neu/HER2) [16], ErbB3 (HER3) [17], and ErbB4 (HER4) [18]. We initially addressed the issue of whether the EGF family and its receptor family were expressed in A7r5
Discussion
In the present study, we demonstrated that amphiregulin, epiregulin, heregulin, and HB-EGF are expressed as evidenced by RT-PCR analysis. The EGF family can be divided into two subgroups on the basis of their direct binding to specific receptors [20], [21]. Members of the first group, which includes amphiregulin, epiregulin, HB-EGF, EGF, TGF-α, and betacellulin, bind directly to the classic EGF receptor. Members of the other group, which includes heregulin, bind directly to the ErbB3 and ErbB4.
Acknowledgements
We are grateful to Dr. Takio Hayashi for encouragement and Mrs. Mari Kurata for excellent technical assistance.
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Endothelin-1 (ET-1) increases the expression of remodeling genes in vascular smooth muscle through linked calcium and cAMP pathways: Role of a phospholipase A<inf>2</inf>(cPLA<inf>2</inf>)/cyclooxygenase-2 (COX-2)/prostacyclin receptor-dependent autocrine loop
2010, Journal of Biological ChemistryCitation Excerpt :Screening of both probe sets against the CRE arrays revealed that at least five gene transcripts were up-regulated by ET-1 namely, amphiregulin, follistatin, COX-2, inhibin-β-A, and CYR61 (Fig. 5A), ET-1 induced increases in mRNA was confirmed by Q-PCR against cDNA synthesized from total RNA extracted from HPASMC treated with ET-1 for various times (Fig. 5, A–F). Amphiregulin is a member of the EGF family of growth factors (32) a potent smooth muscle mitogen (33) that can undergo transcriptional co-induction with other members of the EGF family (34). To determine whether other members of the EGF family were also induced by ET-1 we designed PCR primers for EGF, transforming growth factor α, betacellulin, epiregulin, and HB-EGF and performed Q-PCR.
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Also corresponding author.