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Functional and molecular evidence for expression of renin angiotensin system in COS‐7 cells
Author(s) -
Chodavarapu Harshita,
Elased Khalid M,
DiFulvio Mauricio
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.1088.1
Subject(s) - renin–angiotensin system , western blot , microbiology and biotechnology , blot , immunohistochemistry , cell culture , angiotensin ii , angiotensin converting enzyme 2 , lysis , angiotensin ii receptor type 1 , biology , chemistry , receptor , biochemistry , endocrinology , medicine , gene , immunology , blood pressure , genetics , disease , covid-19 , infectious disease (medical specialty)
The renin angiotensin system (RAS) is a major regulator of renal and cardiovascular functions. There is a need for a rapid throughput assay in transfectable cell culture to study the cross talk between different arms of the RAS. COS‐7 is an easily transfectable cell line derived from the kidney of the African Green Monkey, Cercopithecus aethiops. Previous studies indicated that COS‐7 cell lacks angiotensin type I receptor (AT1R). The aim of this study was to investigate whether COS‐7 cells endogenously express RAS components. Cells were grown to 90% confluency and homogenized in lysis buffer containing protease inhibitors. Western blot and immunohistochemistry were performed to examine the RAS protein expression. ACE and ACE2 activities were measured. RT‐PCR and specific sets of primers were used to amplify ACE, ACE2, AT1 and renin transcripts. Western blot analysis demonstrated expression of AT1 (43 kDa), ACE (195 kDa), ACE2 (90 kDa) and renin (41 kDa) at the protein level. Further, immunohistochemistry experiments also demonstrated expression of ACE, ACE2, AT1 and renin in COS7. At the functional level, ACE and ACE2 activities in COS‐7 lysate were selectively inhibited by lisinopril and MLN‐4760 respectively. Conclusion This is the first report to demonstrate functional endogenous expression of the RAS in COS‐7 cells.