Effect of antisense RNA targeting Polo-like kinase 1 on cell growth in A549 lung cancer cells

J Huazhong Univ Sci Technolog Med Sci. 2008 Feb;28(1):22-6. doi: 10.1007/s11596-008-0106-9.

Abstract

In order to investigate the effect of Polo-like kinase-1 (Plk1) depletion on cell cycle progression and cell growth in lung cancer cells, a recombinant plasmid containing antisense RNA targeting Plk1 (pcDNA3-Plk1) was transfected into A549 cells by lipofectine. RT-PCR and Western-blot were used to detect the Plk1 gene expression. Cell proliferation was evaluated by direct cell counting and bromodeoxyuridine (BrdU) labeling. Cell cycle distribution and apoptosis were examined by flow cytometry, and the inhibition rate (IR) by vinorebline (NVB) was determined by MTT assay. The results showed that after transfection of pcDNA3-Plk1 into A549 cells, the expression levels of Plk1 mRNA and protein were greatly decreased. In pcDNA3-Plk1 transfected groups, abnormal morphological changes of cells and growth inhibition were observed, and the BrdU labeling index was significantly lower than in the control groups (P<0.05). Cells in pcDNA3-Plk1 transfected groups were arresed in G2/M phase and apoptosis was detectable 72 h post transfection. IR induced by vinorebline in pcDNA3-Plk1 transfected groups was significantly higher than in other groups. These data suggested that antisense RNA targeting Plk1 could suppress the Plk1 expression, and therefore, significantly inhibit cell proliferation and induce cell cycle arrest and apoptosis. Moreover, it sensitized lung cancer cells to chemotherapy.

MeSH terms

  • Apoptosis
  • Cell Cycle
  • Cell Cycle Proteins / genetics*
  • Cell Line, Tumor
  • Cell Proliferation*
  • Gene Expression Regulation, Neoplastic*
  • Genetic Techniques
  • Genetic Therapy / methods*
  • Genetic Vectors
  • Humans
  • Lung Neoplasms / enzymology
  • Lung Neoplasms / genetics*
  • Models, Genetic
  • Protein-Serine-Threonine Kinases / genetics*
  • Proto-Oncogene Proteins / genetics*
  • RNA, Antisense / genetics*
  • Time Factors
  • Transfection

Substances

  • Cell Cycle Proteins
  • Proto-Oncogene Proteins
  • RNA, Antisense
  • Protein-Serine-Threonine Kinases
  • polo-like kinase 1