Mobile website
Basic Research
expression in lung adenocarcinoma cell line A549 and its inhibiory effect on cell growth
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective:To investigate the inhibitory effect of inhibitor of differentiation 3 (Id3) on growth of human lung adenocarcinoma cell line A549. Methods: Recombinant eukaryotic expression vector pEGFP/Id3 was constructed and transfected into A549 cells by liposomemediated method. Expression of pEGFP/Id3 in A549 cells was analyzed by flow cytometry (FCM), fluorescence microscopy, semiquantitative RTPCR and immunocytochemistry. The growth inhibitory rate of A549 cells was examined by MTT assay; cell cycle change was evaluated by PI (propidium iodide) staining method. Cell apoptotic rate and nuclear morphology were detected by Annexin V/7AAD and Hoechst33258 staining. Results: The recombinant eukaryotic expression vector pEGFP/Id3 was successfully constructed. The expression of EGFP reached the peak 4872 h after transfection; the expresion of pEGFPtransfected group was higher than that of the pEGFP/Id3 group. RTPCR and immunocytochemistry staining showed that Id3 mRNA and protein were effectively expressed in pEGFP/Id3transfected A549 cells. The growth of cells in pEGFP/Id3 transfected cells was significantly inhibited 4872 h after transfection（P<0.01）. More cells were blocked in G0/G1 phase in pEGFP/Id3transfected group compared with pEGFP group（P<0.05）. Annexin V/7AAD showed that the apoptotic rate of pEGFP/Id3 group （\[10.67±2.60\]%）were significantly higher than those of the control group（\[2.35 ± 0.95\]%）and pEGFP group（\[3.39±2.21\] %）（P<0.05）. Hoechst33258 staining also showed that the cells in pEGFP/Id3 group had typical apoptotic morphology.Conclusion: Exogenous Id3 gene expression in A549 cells can induce cell growth inhibition and apoptosis of A549 cells.
Keywords:
Project Supported:
Supported by the Talent Peak Foundation of Jiangsu province (No. 050203D)
Clc Number:
