Cytotoxicity and Proteomics Analyses of OSU03013 in Lung Cancer

Abstract

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 (1f4 AM) cytotoxicity in all lung cancer cell lines tested 48 hours posttreatment. OSU03013 caused cell cycle G1phase arrest and showed phosphatidylserine early apoptosis via endoplasmic reticulum stress. Several proteins such as heat shock protein 27, 70, and 90, CDC2, a-tubulin, annexin A3, cAMP-dependent protein kinase, glycogen synthase kinase 3-beta, and h-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 h-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.

Quantitative proteome analysis reveals annexin A3 as a novel biomarker in lung adenocarcinoma

“…It has been reported that ANXA3 could promote angiogenesis, possibly through stimulating VEGF and bFGF production, suggesting that ANXA3 might play critical roles in cancer metastasis via promoting angiogenesis 50. Recent studies have also shown that: (a) ANXA3 is necessary for DNA replication in cultured hepatocytes 51; ANXA3 abundance is increased in approximately two‐thirds of colorectal tumours 52; ANXA3 is down‐regulated in prostate cancer cells and an independent adverse prognostic factor 53; and OSU03 013, a derivative of celecoxib, exhibits its antitumour activity by decreasing ANXA3 expression in A549, CL1‐1 and H1435 lung cancer cell lines 54. Our results, together with previous reports, strongly suggest that ANXA3 might serve as a novel biomarker for lymph node metastasis and prognosis in lung AdC, and play an important role in lung AdC progression.…”

Proteomic Analysis of Celecoxib on Chondrocytes from Patients with Osteoarthritis

“…Several comprehensive proteomic analyses of cancer cells treated with celecoxib have been studied in lung can- cer, oral squamous cell carcinoma and colorectal carcinoma [22]- [24]. Another study used cardiomyocytes to investigate the cardiovascular toxicity of NSAIDs including celecoxib [25].…”

Dual Functions of the C5a Receptor as a Connector for the K562 Erythroblast-Like Cell-THP-1 Macrophage-Like Cell Island and as a Sensor for the Differentiation of the K562 Erythroblast-Like Cell during Haemin-Induced Erythropoiesis

“…The K562 cells were differentiated into K562 basophilic-EB-like cells using 30  μ M haemin, and the monocytic THP-1 cells were differentiated into THP-1 macrophage-like cells using 160 nM phorbol-12-myristate-13-acetate (PMA) (Sigma) [21]. To select for apoptosis-resistant clones (EGFP-ANXA3 K562 R cells) using limiting dilution methods, the EGFP-ANXA3 K562 cells were cultured for 3 days in the presence of 1 mM MnCl 2 [18, 22]. To examine the haemin-induced differentiation of the K562 cells using fluorescence-activated cell sorting (FACS), the cells were stained with FITC-conjugated anti-CD71 and PE-conjugated anti-CD235a mouse IgGs for 30 min at 4°C.…”