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Combined Cyclin D1 and Bcl-xL Gene Therapy Attenuated Cell Growth and Promoted Cell Apoptosis in Non-small-cell Lung Cancer

Author: ChenYing
Tutor: XiongWeiNing
School: Huazhong University of Science and Technology
Course: Internal Medicine
Keywords: Lung cancer RNA Interference B-cell lymphoma-xl Cyclin D1 Apoptosis
CLC: R734.2
Type: Master's thesis
Year: 2011
Downloads: 39
Quote: 0
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Abstract


Objectives: To determine combined Cyclin D1 and Bcl-xL’s potential feasibility as a therapeutic target, we constructed a recombinant plasmid pcDNA6.2-GW/EmGFP-miR that expressed a CMV promoter-driven microRNA targeting Cyclin D1 (Cyclin D1 miRNA), Bcl-xL (Bcl-xL miRNA) and these two genes (Cyclin D1-Bcl-xL miRNA). Then the vector’s ability to induce RNA interference in vitro and alter apoptosis induction in lung cancer was assessed.Methods: Four groups was used: the group of blank control (untreated cells), Cyclin D1 miRNA, Bcl-xL miRNA and Cyclin D1-Bcl-xL miRNA (transfected cells). The cell lines were used A549 and NCI-H441. The levels of mRNA and protein of Bcl-xL or Cylin D1 were detected by RT-PCR and Western blot. Apoptosis and proliferation of lung cancer cells were evaluated by dimethylthiazol-diphenyltetrazolium bromide (MTT), cell count and flow cytometry.Results: Our results suggested that the recombinant plasmid pcDNA6.2-GW/EmGFP-miR can adequately mediated RNAi in A549 and NCI-H441 cells. The expressions’levels of mRNA and protein in the groups Cyclin D1 miRNA, Bcl-xL miRNA and Cyclin D1-Bcl-xL miRNA (stable transfectants) were significantly reduced compared to untreated cells (P<0.05). There was no statistical difference in the three interventional groups (P>0.05). The cell inhibition rate and apoptosis in the three groups were increased (P<0.05). Combined interference of two genes had higher cell inhibition rate and cell apoptosis than a single one (P<0.05).Conclusions: The molecular targeting in repression of Cyclin D1 and Bcl-xL genes in combination as a therapeutic strategy for human malignancies.

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CLC: > Medicine, health > Oncology > Respiratory system tumors > Lung tumors
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