Presence of angiotensinogen messenger RNA in various cultured cell lines

Valerie Fatigati, Jacqueline M. Washington, Kevin R. Lynch, Michael J. Peach, Edward D. Miller

Research output: Contribution to journalArticle

Abstract

The presence of angiotensinogen messenger RNA (mRNA) was assessed in total RNA extracted from hepatoma, glioma, neuroblastoma, and glioma-neuroblastoma hybrid cell lines. Total RNA from 1 × 107 cells was extracted, transferred to a membrane, and hybridized with a 32P-labeled, full-length (1650-base pair) rat angiotensinogen complementary DNA (cDNA). Angiotensinogen RNA sequences could be definitively detected only in hepatoma cells. Steroids were used in an attempt to increase the angiotensinogen mRNA level. Dexamethasone (2 × 10-6M) or 17 β-estradiol (1 × 10 -7 M) was added to the cultures 18 to 24 hours prior to harvest. Dexamethasone treatment of the hepatoma cells resulted in a large increase in angiotensinogen mRNA, whereas estradiol had no effect. Steroids failed to induce detectable levels of angiotensinogen mRNA in total RNA from the other cell lines. That the RNA was intact was ensured by hybridizing duplicate Northern blots to a 32P-Iabeled actin cDNA. Actin mRNA sequences were detected in all cell lines. Blot hybridization of poly (A)+ RNA resulted in the visualization of a weak angiotensinogen mRNA signal for a glioma cell line and a glioma-neuroblastoma hybrid line. However, the ability to detect angiotensinogen mRNA in a cell may depend on the phenotype expressed, which can be governed by culture conditions.

Original languageEnglish (US)
Pages (from-to)III-25-III-30
JournalHypertension
Volume9
Issue number6, Part 2
DOIs
StatePublished - Jun 1987

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Keywords

  • Angiotensinogen
  • Complementary DNA
  • Cultured cells
  • Messenger RNA

ASJC Scopus subject areas

  • Internal Medicine

Cite this

Fatigati, V., Washington, J. M., Lynch, K. R., Peach, M. J., & Miller, E. D. (1987). Presence of angiotensinogen messenger RNA in various cultured cell lines. Hypertension, 9(6, Part 2), III-25-III-30. https://doi.org/10.1161/01.hyp.9.6_pt_2.iii25