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  • Title: [Effect of antisense RNA targeting Polo-like kinase 1 on cell cycle of lung cancer cell line A549].
    Author: Zhou Q, Bai M, Su Y.
    Journal: Ai Zheng; 2005 Feb; 24(2):149-54. PubMed ID: 15694023.
    Abstract:
    BACKGROUND & OBJECTIVE: Expression of polo-like kinase 1 (Plk1), which has several functions in mitotic progression, is elevated in lung adenocarcinoma cell line A549. It suggests that over-expression of Plk1 relates to chemotherapy- and radiotherapy-resistance, and poor prognosis of lung cancer patients. This study was to investigate the effect of down-regulation of Plk1 on cell cycle of A549 cells using the technique of antisense RNA. METHODS: Antisense RNA targeting Plk1 (pcDNA3.0-Plk1, pc3.0P) was designed and constructed, and transfected into A549 cells. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to examine expression of Plk1 gene. Cell proliferation was evaluated by BrdU stain; cell cycle changes and apoptosis were examined by flow cytometry; expression of alpha-tubulin was detected by immunofluorescence. RESULTS: mRNA level of Plk1 was significantly lower in pc3.0P-transfected A549 cells than in control cells (P < 0.05). pc3.0P reduced Plk1 mRNA in A549 cells by 46.75% 24 h after transfection, and by 61.84% 48 h after transfection. Protein level of Plk1 was also decreased after transfection. Positive rate of BrdU stain in experimental cells was only 25.59% 48 h after transfection, which was significantly lower than that in control cells (P < 0.05). A549 cells showed a strong G(2)/M arrest and apoptosis 72 h after transfection. alpha-tubulin staining showed absence of microtubule connection,and spindle abnormalities. CONCLUSION: Down-regulation of Plk1 interferes spindle formation, and inhibits cell proliferation, induces cell cycle arrest and apoptosis.
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