Use of recombinant calpain-2 siRNA adenovirus to assess calpain-2 modulation of lung endothelial cell migration and proliferation

Kai Qiu, Yunchao Su, Edward R. Block

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

In this study, we developed an adenoviral vector harboring calpain-2 siRNA expression unit in which sense and anti-sense strands composing the siRNA duplex were connected by a loop and transcribed into a siRNA in porcine pulmonary artery endothelial cells (PAEC). We screened one efficient adenoviral vector Ad/si-m187 and found that Ad/si-m187 successfully exerted a gene knockdown effect on calpain-2 mRNA transcription and protein expression levels. The protein content of calpain-2 was reduced by 30%-80% in PAEC infected with Ad/si-m187 in comparison to a control adenoviral vector Ad/si-luc. The mRNA levels of calpain-2 were measured by real-time PCR and were decreased by 60%-100% and in a dose dependent manner. In correspondence to silencing calpain-2 gene expression, calpain-2 activity was decreased significantly. We further evaluated the role of calpain-2 in endothelial cell migration and proliferation. PAEC infected with Ad/si-m187 displayed impaired migration and cell proliferation in comparison to cells infected with control adenoviral vector (Ad/si-luc). These results indicate that adenoviral vector harboring calpain-2 siRNA expression unit is a valuable tool to study the biology of calpains and that calpain-2 plays an important role in lung endothelial cell migration and proliferation.

Original languageEnglish (US)
Pages (from-to)69-78
Number of pages10
JournalMolecular and Cellular Biochemistry
Volume292
Issue number1-2
DOIs
StatePublished - Nov 1 2006
Externally publishedYes

Fingerprint

Calpain
Endothelial cells
Adenoviridae
Small Interfering RNA
Cell Movement
Endothelial Cells
Cell Proliferation
Modulation
Lung
Pulmonary Artery
Gene Knockdown Techniques
Messenger RNA
Cell proliferation
Transcription
Gene expression
Real-Time Polymerase Chain Reaction
Proteins
Swine
Genes
Gene Expression

Keywords

  • Adenovirus
  • Angiogenesis
  • Calpains
  • Endothelium
  • SiRNA

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Use of recombinant calpain-2 siRNA adenovirus to assess calpain-2 modulation of lung endothelial cell migration and proliferation. / Qiu, Kai; Su, Yunchao; Block, Edward R.

In: Molecular and Cellular Biochemistry, Vol. 292, No. 1-2, 01.11.2006, p. 69-78.

Research output: Contribution to journalArticle

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