Synergistic Effect of Sulindac and Simvastatin on Apoptosis in Lung Cancer A549 Cells through AKT-Dependent Downregulation of Survivin
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Purpose Non-steroidal anti-inflammatory drugs (NSAIDs) and statins are potential chemopreventiveor chemotherapeutic agents. The mechanism underlying the deregulation of survivin byNSAIDs and statins in human non-small cell lung cancer cells has not been elucidated. Inthis study, we investigated the synergistic interaction of sulindac and simvastatin in lungcancer A549 cells. Materials and MethodsCell viability was measured by an MTT assay, while the expression of apoptotic markers,AKT, and survivin in response to sulindac and simvastatin was examined by Westernblotting. DNA fragmentation by apoptosis was analyzed by flow cytometry in A549 cells. Reactive oxygen species (ROS) generation was measured by flow cytometry using H2DCFDAand MitoSOX Red, and the effects of pretreatment with N-acetylcysteine were tested. Theeffects of AKT on survivin expression in sulindac- and simvastatin-treated cells wereassessed. Survivin was knocked down or overexpressed to determine its role in apoptosisinduced by sulindac and simvastatin. ResultsSulindac and simvastatin synergistically augmented apoptotic activity and intracellular ROSproduction in A549 cells. Inhibition of AKT by siRNA or LY294002 inhibited survivin, whileAKT overexpression markedly increased survivin expression, even in the presence ofsulindac and simvastatin. Moreover, survivin siRNA enhanced sulindac- and simvastatininducedapoptosis. In contrast, survivin upregulation protected against sulindac- andsimvastatin-induced apoptosis. ConclusionCombined treatment with sulindac and simvastatin augmented their apoptotic potential inlung cancer cells through AKT signaling-dependent downregulation of survivin. These resultsindicate that sulindac and simvastatin may be clinically promising therapies for the preventionof lung cancer.
Abstract
Purpose Non-steroidal anti-inflammatory drugs (NSAIDs) and statins are potential chemopreventiveor chemotherapeutic agents. The mechanism underlying the deregulation of survivin byNSAIDs and statins in human non-small cell lung cancer cells has not been elucidated. Inthis study, we investigated the synergistic interaction of sulindac and simvastatin in lungcancer A549 cells. Materials and MethodsCell viability was measured by an MTT assay, while the expression of apoptotic markers,AKT, and survivin in response to sulindac and simvastatin was examined by Westernblotting. DNA fragmentation by apoptosis was analyzed by flow cytometry in A549 cells. Reactive oxygen species (ROS) generation was measured by flow cytometry using H2DCFDAand MitoSOX Red, and the effects of pretreatment with N-acetylcysteine were tested. Theeffects of AKT on survivin expression in sulindac- and simvastatin-treated cells wereassessed. Survivin was knocked down or overexpressed to determine its role in apoptosisinduced by sulindac and simvastatin. ResultsSulindac and simvastatin synergistically augmented apoptotic activity and intracellular ROSproduction in A549 cells. Inhibition of AKT by siRNA or LY294002 inhibited survivin, whileAKT overexpression markedly increased survivin expression, even in the presence ofsulindac and simvastatin. Moreover, survivin siRNA enhanced sulindac- and simvastatininducedapoptosis. In contrast, survivin upregulation protected against sulindac- andsimvastatin-induced apoptosis. ConclusionCombined treatment with sulindac and simvastatin augmented their apoptotic potential inlung cancer cells through AKT signaling-dependent downregulation of survivin. These resultsindicate that sulindac and simvastatin may be clinically promising therapies for the preventionof lung cancer.
참고문헌 (25)
- Omenn GS, "Chemoprevention of lung cancer is proving difficult and frustrating, requiring new approaches", J Natl Cancer Inst, 92, pp.959~960 (2000)
- Narayanan BA, "Regression of mouse prostatic intraepithelial neoplasia by nonsteroidal anti-inflammatory drugs in the transgenic adenocarcinoma mouse prostate model", Clin Cancer Res, 10, pp.7727~7737 (2004)
- Ulrich CM, "Non-steroidal anti-inflammatory drugs for cancer prevention : promise, perils and pharmacogenetics", Nat Rev Cancer, 6, pp.130~140 (2006)
- Albayrak A, "Gastric anti-ulcerative and anti-inflammatory activity of metyrosine in rats", Pharmacol Rep, 62, pp.113~119 (2010)
- Krysan K, "The potential and rationale for COX-2 inhibitors in lung cancer", Anticancer Agents Med Chem, 6, pp.209~220 (2006)
- Solomon SD, "Cardiovascular risk associated with celecoxib in a clinical trial for colorectal adenoma prevention", N Engl J Med, 352, pp.1071~1080 (2005)
- Psaty BM, "Risks and benefits of celecoxib to prevent recurrent adenomas", N Engl J Med, 355, pp.950~952 (2006)
- Goldstein JL, "Regulation of the mevalonate pathway", Nature, 343, pp.425~430 (1990)
- Chan KK, "The statins as anticancer agents", Clin Cancer Res, 9, pp.10~19 (2003)
- Demierre MF, "Statins and cancer prevention", Nat Rev Cancer, 5, pp.930~942 (2005)
- Cafforio P, "Statins activate the mitochondrial pathway of apoptosis in human lymphoblasts and myeloma cells", Carcinogenesis, 26, pp.883~891 (2005)
- Jalving M, "Review article : the potential of combinational regimen with non-steroidal anti-inflammatory drugs in the chemoprevention of colorectal cancer", Aliment Pharmacol Ther, 21, pp.321~339 (2005)
- Hwang KE, "Synergistic induction of apoptosis by sulindac and simvastatin in A549 human lung cancer cells via reactive oxygen speciesdependent mitochondrial dysfunction", Int J Oncol, 43, pp.262~270 (2013)
- Crissman HA, "Cell cycle-related changes in chromatin structure detected by flow cytometry using multiple DNA fluorochromes", Eur J Histochem, 37, pp.129~138 (1993)
- Elbashir SM, "Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells", Nature, 411, pp.494~498 (2001)
- Tamm I, "IAP-family protein survivin inhibits caspase activity and apoptosis induced by Fas(CD95), Bax, caspases, and anticancer drugs", Cancer Res, 58, pp.5315~5320 (1998)
- Krajewska M, "Elevated expression of inhibitor of apoptosis proteins in prostate cancer", Clin Cancer Res, 9, pp.4914~4925 (2003)
- Minami T, "Sulindac enhances the proteasome inhibitor bortezomib-mediated oxidative stress and anticancer activity", Clin Cancer Res, 11, pp.5248~5256 (2005)
- Mukhopadhyay P, "Simple quantitative detection of mitochondrial superoxide production in live cells", Biochem Biophys Res Commun, 358, pp.203~208 (2007)
- Li F, "Control of apoptosis and mitotic spindle checkpoint by survivin", Nature, 396, pp.580~584 (1998)
- Kaneko R, "Survivin down-regulation plays a crucial role in 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitorinduced apoptosis in cancer", J Biol Chem, 282, pp.19273~19281 (2007)
- Ohashi H, "Phosphatidylinositol 3-kinase", Akt regulates angiotensin II-induced inhibition of apoptosis in microvascular endothelial cells by governing survivin expression and suppression of caspase-3 activity, 2004, pp.94, 785~793
- Fornaro M, "Fibronectin protects prostate cancer cells from tumor necrosis factor-alpha-induced apoptosis via the AKT", survivin pathway, 2003, pp.278, 50402~50411
- Vaira V, "Regulation of survivin expression by IGF-1", mTOR signaling, 2007, pp.26, 2678~2684
- You L, "Inhibition of Wnt-2-mediated signaling induces programmed cell death in non-small-cell lung cancer cells", Oncogene, 23, pp.6170~6174 (2004)