Inhibition of Invasiveness of Human Lung Cancer Cells by Adenovirus-mediated Gene Transfer of Antisense RNA for Urokinase Receptor

Xing Hui Sun, Li Tan, Ping Li, Yu Qing Zhang, Xia Wang, Min Hou, Hou Yan Song, Yun Song Zhu

Research output: Contribution to journalArticle

Abstract

The effect of antisense RNA for urokinase receptor (uPAR) on inhibition of invasiveness of human lung giant cancer cell lines 95D was observed. 500bp fragment of uPAR cDNA between -46bp ∼ +454bp was amplified, and recombined into plasmid pAdeno-X. The recombinant vector was named pAdeno-X-uPAR (-) and pAdeno-X-uPAR (+) respectively. 7 days after transfecting HEK293 cell with linearized pAdeno-X-uPAR (-) and pAdeno-X-uPAR (+), the recombinant adenovirus can be obtained, which were named Ad-uPAR (-) and Ad-uPAR (+) respectively. The virus titre (pfu/ml) of Ad-uPAR (-) was 1. 5 × 108, and the virus titre of Ad-uPAR (+) was 0.5 × 108. 95D cells were infected with Ad-uPAR (-) and Ad-uPAR (+) in different multiplicity of infection (MOI). Norther blot analysis could detected the expression of antisense and sense RNA for 500 bp fragment of uPAR gene. With the increase of MOI, Western blot analysis indicated that with AD-uPAR (-) infection the protein level of uPAR of 95D cells decreased, and modified Boyden's Chamber assay suggested that the invasive ability of 95D cells also decreased obviously. In 95D cells infected with Ad-uPAR (+), both the mRNA and protein level of uPAR did not decrease, and cells still had high invasive ability. The results indicated that adenovirus is an efficient vector for transferring antisense RNA for uPAR into cells, and antisense RNA for uPAR could obviously inhibit the invasive ability of 95D human lung cancer cells.

Original languageEnglish (US)
Pages (from-to)761-766
Number of pages6
JournalProgress in Biochemistry and Biophysics
Volume30
Issue number5
StatePublished - Oct 1 2003

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Gene transfer
Antisense RNA
Urokinase-Type Plasminogen Activator
Transfer RNA
Adenoviridae
Lung Neoplasms
Cells
Viruses
Genes
Viral Load
Assays
Proteins
Plasmids
Complementary DNA
RNA
Infection
Messenger RNA
HEK293 Cells
Giant Cells
Western Blotting

Keywords

  • Adenovirus
  • Antisense gene
  • Cancer cells
  • Invasiveness
  • Urokinase receptor

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry

Cite this

Inhibition of Invasiveness of Human Lung Cancer Cells by Adenovirus-mediated Gene Transfer of Antisense RNA for Urokinase Receptor. / Sun, Xing Hui; Tan, Li; Li, Ping; Zhang, Yu Qing; Wang, Xia; Hou, Min; Song, Hou Yan; Zhu, Yun Song.

In: Progress in Biochemistry and Biophysics, Vol. 30, No. 5, 01.10.2003, p. 761-766.

Research output: Contribution to journalArticle

Sun, Xing Hui ; Tan, Li ; Li, Ping ; Zhang, Yu Qing ; Wang, Xia ; Hou, Min ; Song, Hou Yan ; Zhu, Yun Song. / Inhibition of Invasiveness of Human Lung Cancer Cells by Adenovirus-mediated Gene Transfer of Antisense RNA for Urokinase Receptor. In: Progress in Biochemistry and Biophysics. 2003 ; Vol. 30, No. 5. pp. 761-766.
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AU - Zhang, Yu Qing

AU - Wang, Xia

AU - Hou, Min

AU - Song, Hou Yan

AU - Zhu, Yun Song

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AB - The effect of antisense RNA for urokinase receptor (uPAR) on inhibition of invasiveness of human lung giant cancer cell lines 95D was observed. 500bp fragment of uPAR cDNA between -46bp ∼ +454bp was amplified, and recombined into plasmid pAdeno-X. The recombinant vector was named pAdeno-X-uPAR (-) and pAdeno-X-uPAR (+) respectively. 7 days after transfecting HEK293 cell with linearized pAdeno-X-uPAR (-) and pAdeno-X-uPAR (+), the recombinant adenovirus can be obtained, which were named Ad-uPAR (-) and Ad-uPAR (+) respectively. The virus titre (pfu/ml) of Ad-uPAR (-) was 1. 5 × 108, and the virus titre of Ad-uPAR (+) was 0.5 × 108. 95D cells were infected with Ad-uPAR (-) and Ad-uPAR (+) in different multiplicity of infection (MOI). Norther blot analysis could detected the expression of antisense and sense RNA for 500 bp fragment of uPAR gene. With the increase of MOI, Western blot analysis indicated that with AD-uPAR (-) infection the protein level of uPAR of 95D cells decreased, and modified Boyden's Chamber assay suggested that the invasive ability of 95D cells also decreased obviously. In 95D cells infected with Ad-uPAR (+), both the mRNA and protein level of uPAR did not decrease, and cells still had high invasive ability. The results indicated that adenovirus is an efficient vector for transferring antisense RNA for uPAR into cells, and antisense RNA for uPAR could obviously inhibit the invasive ability of 95D human lung cancer cells.

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