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Overexpression of Id3 induces apoptosis of A549 human lung adenocarcinoma cells.
MedLine Citation:
PMID:  22151756     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Objectives:  Inhibitor of differentiation 3 (Id3) protein has been implicated in the control of multiple cell death signalling pathways and in aetiology of numerous diseases. The aims of this study were to construct a recombinant eukaryotic expression vector (pEGFP/Id3), containing human Id3 (hId3) fused with enhanced green fluorescent protein (EGFP), and to determine effects of ectopic Id3 overexpression, on human lung adenocarcinoma cell (A549) proliferation. Materials and methods:  Human Id3 cDNA was inserted into pEGFP-N1 vector to yield the recombinant eukaryotic expression vector pEGFP/Id3. Cells were transfected with pEGFP or pEGFP/Id3, and proliferation of EGFP-expressing cells was monitored by flow cytometry (FCM) and confocal fluorescence microscopy. RT-PCR, immunoblotting and immunocytochemistry were used to assess Id3 mRNA transcription and protein expression. Apoptosis was evaluated by Annexin V/7-AAD staining and FCM, while nuclear morphology of apoptotic cells was examined using Hoechst 33258 staining. Results:  Over 4 days transfection with pEGFP, the proportion of EGFP-positive A549 cells peaked at approximately 60% by 48 h and remained stable over the next 48 h. In contrast, the proportion of EGFP-positive cells in cultures transfected with pEGFP/Id3 decreased from a peak of 60% at 48 h to <5% at 96 h, suggesting that Id3 expression inhibited cell proliferation or survival. Annexin V/7-AAD and Hoechst 33258 staining revealed significantly higher rates of apoptosis in pEGFP/Id3-transfected cells. Conclusion:  Overexpression of Id3 triggered apoptosis in A549 human lung adenocarcinoma cells, implicating Id3 in negative control of tumour growth. These Id3-induced pro-apoptotic signalling pathways require further study, but this preliminary investigation suggests that Id3 regulation could be exploited in anti-tumour therapies.
Authors:
X-J Li; C-D Zhu; W Yu; P Wang; F-F Chen; X-Y Xia; B Luo
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-12-7
Journal Detail:
Title:  Cell proliferation     Volume:  -     ISSN:  1365-2184     ISO Abbreviation:  -     Publication Date:  2011 Dec 
Date Detail:
Created Date:  2011-12-13     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9105195     Medline TA:  Cell Prolif     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
© 2011 Blackwell Publishing Ltd.
Affiliation:
Center of Clinical Laboratory Science, Jinling Hospital, School of Medicine, Nanjing University, Nanjing, China.
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