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Altered protein-expressing profile in hPNAS4-induced apoptosis in A549 human lung adenocarcinoma cells
Lan-Tu Gou1, Ai-Ping Tong, Fei Yan
1State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041, Sichuan Province, China.
Abstract:
Human PNAS4 (hPNAS4) is a recently identified pro-apoptosis gene, which is able to induce apoptosis in A549 human lung adenocarcinoma cells following its overexpression. In this work, we investigated the changes of protein profile in hPNAS4-induced apoptosis in A549 cells through proteomic strategy consisting of two-dimensional electrophoresis (2-DE) coupled with MALDI-Q-TOF mass spectrometry. A total of 20 different proteins with more than 3.0-fold change in expression, including 5 up-regulated and 15 down-regulated proteins were successfully identified by database search. The mRNA transcription levels of the different proteins were further examined by RT-PCT. Functional analyses showed these different proteins are involved in diverse biological processes including metabolism, proteolysis, signal transduction, apoptosis, and redox regulation. Two essential apoptosis-associated protein, annexin A1 and prothymosin alpha, were confirmed by Western blot and showed consistent changes with proteomic detection. Our data provide molecular evidence and possible associated pathway in hPNAS4-induced apoptosis through proteomic strategy, which should be contributed to further investigation on biological function of hPNAS4.
Insights
Human PNAS4 (hPNAS4) induces apoptosis in lung cancer cells. Proteomics identified 20 proteins involved in cell death pathways, offering insights into hPNAS4
Area of Science:
- Molecular Biology
- Cancer Research
- Proteomics
Background:
- Human PNAS4 (hPNAS4) is a newly discovered pro-apoptosis gene.
- Overexpression of hPNAS4 induces apoptosis in A549 human lung adenocarcinoma cells.
Purpose of the Study:
- To investigate protein profile changes during hPNAS4-induced apoptosis in A549 cells.
- To identify proteins and pathways associated with hPNAS4-mediated apoptosis.
Main Methods:
- Proteomic strategy: two-dimensional electrophoresis (2-DE) coupled with MALDI-Q-TOF mass spectrometry.
- Analysis of mRNA transcription levels using RT-PCR.
- Confirmation of protein changes via Western blot.
Main Results:
- Identified 20 differentially expressed proteins (5 up-regulated, 15 down-regulated) with >3.0-fold change.
- These proteins are involved in metabolism, proteolysis, signal transduction, apoptosis, and redox regulation.
- Annexin A1 and prothymosin alpha, key apoptosis proteins, showed consistent changes.
Conclusions:
- Proteomic analysis provides molecular evidence for hPNAS4-induced apoptosis.
- Identified proteins and pathways offer insights into the biological function of hPNAS4.
- Findings contribute to further research on hPNAS4 and its role in cancer cell death.
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