Identification and characterization of functional angiotensin II type 1 receptors on immortalized human fetal aortic vascular smooth muscle cells

Molecular and Cellular Endocrinology
M M MartinT S Elton

Abstract

Studies investigating the mechanisms that govern the expression of the human angiotensin II type 1 receptor (hAT(1)R) gene have progressed slowly due to the lack of human cell lines that express the AT(1)R. Recently, however, an immortalized human fetal aortic vascular smooth muscle cell line (FLTR) was generated using an amphotropic recombinant retroviral construct containing the E6/E7 open reading frames of the human papillomavirus type 16. Radioligand binding studies were undertaken to determine whether angiotensin II (Ang II) receptors were expressed on these cells. FLTR cell membranes were shown to express high-affinity Ang II receptors having a B(max) value of 324+/-43 fmol/mg protein and a K(d) of 0.36+/-0.1 nM. In both membranes and intact cells, Ang II, Ang III and the selective AT(1)R antagonist, Losartan, all had a high affinity for the receptor, suggesting that FLTR cells express the AT(1)R subtype. The expression of the hAT(1)R was validated by Northern and Western blot and RT-PCR experiments. In intact FLTR cells, Ang II (100 nM) evoked an increase in intracellular calcium ([Ca(2+)](i)) and induced hyperplasia. Additionally, our results demonstrated that FLTR cells were readily transfected, and hAT(1)R promoter lu...Continue Reading

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Citations

Jan 26, 2010·The Journal of Biological Chemistry·Leslie A CaromileDaniel F Bowen-Pope
Apr 9, 2005·Journal of Inflammation·Gary Robert Smith, Sotiris Missailidis
Oct 28, 2004·Biochimica Et Biophysica Acta·Aaron A DuffyTerry S Elton
Aug 5, 2006·Journal of Cellular Biochemistry·Raquel SoaresMónica Botelho
Oct 18, 2003·Circulation Research·Yingbi ZhouMuthu Periasamy

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