Overexpression of the cellular repressor of E1A-stimulated-genes regulate the rat primary VSMCs differentiation in vitro

Ya Ling Han, Cheng Hui Yan, Hai Wei Liu, Ye Hu, Jian Kang, Xiao Zeng Wang, Shaohua Li

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

To elucidate the role of cellular repressor of E1A-stimulated genes (CREG) in the phenotype modulation of vascular smooth muscle cells (VSMCs), human CREG (hCREG) was overexpressed in primary rat VSMCs was using a mammalian expression vector pRC/CMV-hCREG. Transient transfection of VSMCs with hCREG inhibited cell proliferation and DNA synthesis as evidenced by decrease in BrdU incorporation. Moreover, CREG overexpression increased SM α-actin protein level in VSMCs. Co-transfection of VSMCs with SM α-actin promoter reporter gene with hCREG enhanced SM α-actin promoter activity, suggesting that elevated SM α-actin protein production resulted from increased gene transcription. The mechanism by which CREG regulates VSMC phenotype conversion was further investigated by analysis of interaction of CREG with serum response factor (SRF), an important transcriptional factor involved in VSMC differentiation. CREG co-immunoprecipitated with SRF and CREG overexpression increased the SRF bound to CREG. In addition, gel mobility shift assay and supershift assay showed that CREG bound together with SRF to the CArG site of SM α-actin promoter. These results suggest that CREG acts as a transcriptional factor and interacts with SRF to promote VSMC differentiation.

Original languageEnglish (US)
Pages (from-to)1099-1105
Number of pages7
JournalProgress in Biochemistry and Biophysics
Volume31
Issue number12
StatePublished - Dec 1 2004

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Vascular Smooth Muscle
Smooth Muscle Myocytes
Muscle
Rats
Cell Differentiation
Genes
Serum Response Factor
Actins
Transfection
In Vitro Techniques
Assays
Phenotype
Electrophoretic Mobility Shift Assay
Bromodeoxyuridine
Cell proliferation
Reporter Genes
Transcription
Proteins
Gels
Cell Proliferation

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry

Cite this

Han, Ya Ling ; Yan, Cheng Hui ; Liu, Hai Wei ; Hu, Ye ; Kang, Jian ; Wang, Xiao Zeng ; Li, Shaohua. / Overexpression of the cellular repressor of E1A-stimulated-genes regulate the rat primary VSMCs differentiation in vitro. In: Progress in Biochemistry and Biophysics. 2004 ; Vol. 31, No. 12. pp. 1099-1105.
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abstract = "To elucidate the role of cellular repressor of E1A-stimulated genes (CREG) in the phenotype modulation of vascular smooth muscle cells (VSMCs), human CREG (hCREG) was overexpressed in primary rat VSMCs was using a mammalian expression vector pRC/CMV-hCREG. Transient transfection of VSMCs with hCREG inhibited cell proliferation and DNA synthesis as evidenced by decrease in BrdU incorporation. Moreover, CREG overexpression increased SM α-actin protein level in VSMCs. Co-transfection of VSMCs with SM α-actin promoter reporter gene with hCREG enhanced SM α-actin promoter activity, suggesting that elevated SM α-actin protein production resulted from increased gene transcription. The mechanism by which CREG regulates VSMC phenotype conversion was further investigated by analysis of interaction of CREG with serum response factor (SRF), an important transcriptional factor involved in VSMC differentiation. CREG co-immunoprecipitated with SRF and CREG overexpression increased the SRF bound to CREG. In addition, gel mobility shift assay and supershift assay showed that CREG bound together with SRF to the CArG site of SM α-actin promoter. These results suggest that CREG acts as a transcriptional factor and interacts with SRF to promote VSMC differentiation.",
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Overexpression of the cellular repressor of E1A-stimulated-genes regulate the rat primary VSMCs differentiation in vitro. / Han, Ya Ling; Yan, Cheng Hui; Liu, Hai Wei; Hu, Ye; Kang, Jian; Wang, Xiao Zeng; Li, Shaohua.

In: Progress in Biochemistry and Biophysics, Vol. 31, No. 12, 01.12.2004, p. 1099-1105.

Research output: Contribution to journalArticle

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AU - Yan, Cheng Hui

AU - Liu, Hai Wei

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AU - Kang, Jian

AU - Wang, Xiao Zeng

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