Related Experiment Videos
Cytotoxicity and proteomics analyses of OSU03013 in lung cancer
Yi-Hung Tan1, Kung-Hsueh Lee, Topp Lin
1Department of Life Science, National Taiwan Normal University, Taipei, Taiwan.
Purpose:
Most lung cancer patients have some resistance to and suffer from side effects of conventional chemotherapy. Thus, identification of a novel anticancer drug with better target selectivity for lung cancer treatment is urgently needed.
Experimental Design:
In order to investigate whether OSU03013, a derivative of celecoxib, can be a potential drug for lung cancer treatment, we examined its cytotoxicity mechanisms by flow cytometry and phosphatidylserine staining in A549, CL1-1, and H1435 lung cancer cell lines, which are resistant to the conventional drug, cisplatin. In addition, we identified the affected proteins by proteomics and confirmed the selected proteins by Western blot analysis. We examined the interaction between OSU03013 and potential target protein by molecular modeling.
Results:
Our results indicated that OSU03013 had low-dose (1 approximately 4 microM) cytotoxicity in all lung cancer cell lines tested 48 hours posttreatment. OSU03013 caused cell cycle G1 phase arrest and showed phosphatidylserine early apoptosis via endoplasmic reticulum stress. Several proteins such as heat shock protein 27, 70, and 90, CDC2, alpha-tubulin, annexin A3, cAMP-dependent protein kinase, glycogen synthase kinase 3-beta, and beta-catenin were identified by proteomics and confirmed by Western blot. In addition, molecular modeling showed that OSU03013 competes with ATP to bind to cAMP-dependent protein kinase.
Conclusions:
We identified for the first time that OSU03013 inhibits cAMP-dependent protein kinase activity and causes dephosphorylation of glycogen synthase kinase 3-beta leading to beta-catenin degradation, which is often overexpressed in lung cancer. Our molecular and proteomic results show the potential of OSU03013 as an anticancer drug for lung cancer.
Insights
OSU03013, a novel celecoxib derivative, effectively treats lung cancer by inducing apoptosis and cell cycle arrest. This compound inhibits cAMP-dependent protein kinase, leading to beta-catenin degradation, offering a promising new therapeutic avenue for lung cancer patients.
Area of Science:
- Oncology
- Molecular Biology
- Drug Discovery
Background:
- Lung cancer patients often exhibit resistance to conventional chemotherapy, necessitating novel therapeutic agents.
- Conventional treatments can lead to significant side effects, highlighting the need for targeted therapies.
Purpose of the Study:
- To investigate OSU03013, a celecoxib derivative, as a potential novel drug for lung cancer treatment.
- To elucidate the cytotoxicity mechanisms of OSU03013 in cisplatin-resistant lung cancer cell lines.
Main Methods:
- Cytotoxicity was assessed using flow cytometry and phosphatidylserine staining in A549, CL1-1, and H1435 lung cancer cell lines.
- Proteomics and Western blot analysis were employed to identify and confirm proteins affected by OSU03013.
- Molecular modeling was used to examine the interaction between OSU03013 and its potential target protein.
Main Results:
- OSU03013 demonstrated low-dose cytotoxicity (1-4 microM) in all tested lung cancer cell lines within 48 hours.
- The drug induced G1 phase cell cycle arrest and early apoptosis via endoplasmic reticulum stress.
- Proteomic analysis identified several key proteins, including heat shock proteins, CDC2, alpha-tubulin, annexin A3, cAMP-dependent protein kinase, glycogen synthase kinase 3-beta, and beta-catenin.
Conclusions:
- OSU03013 was identified as an inhibitor of cAMP-dependent protein kinase activity.
- The drug promotes dephosphorylation of glycogen synthase kinase 3-beta, resulting in beta-catenin degradation, a protein frequently overexpressed in lung cancer.
- These findings highlight OSU03013's potential as an effective anticancer drug for lung cancer treatment.