Enhanced apoptosis by pemetrexed and simvastatin in malignant mesothelioma and lung cancer cells by reactive oxygen species-dependent mitochondrial dysfunction and Bim induction

Hwang,Ki-Eun; Kim,Young-Suk; Hwang,Yu-Ri; Kwon,Su-Jin; Park,Do-Sim; Cha,Byong-Ki; Kim,Byoung-Ryun; Yoon,Kwon-Ha; Jeong,Eun-Taik; Kim,Hak-Ryul

doi:10.3892/ijo.2014.2584

Enhanced apoptosis by pemetrexed and simvastatin in malignant mesothelioma and lung cancer cells by reactive oxygen species-dependent mitochondrial dysfunction and Bim induction

Authors:
- Ki-Eun Hwang
- Young-Suk Kim
- Yu-Ri Hwang
- Su-Jin Kwon
- Do-Sim Park
- Byong-Ki Cha
- Byoung-Ryun Kim
- Kwon-Ha Yoon
- Eun-Taik Jeong
- Hak-Ryul Kim
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Affiliations: Department of Internal Medicine, Institute of Wonkwang Medical Science, Wonkwang University School of Medicine, Iksan, Jeonbuk 570-749, Republic of Korea, Department of Laboratory Medicine, Wonkwang University School of Medicine, Iksan, Jeonbuk 570-749, Republic of Korea, Department of Thoracic and Cardiovascular Surgery, Chonbuk National University Medical School, Jeonju, Jeonbuk 561-180, Republic of Korea, Department of Obstetrics and Gynecology, Wonkwang University School of Medicine, Iksan, Jeonbuk 570-749, Republic of Korea, Department of Radiology, Wonkwang University School of Medicine, Iksan, Jeonbuk 570-749, Republic of Korea
Published online on: August 5, 2014 https://doi.org/10.3892/ijo.2014.2584
Pages: 1769-1777

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Abstract

Pemetrexed is a multitarget antifolate currently used for the treatment of malignant mesothelioma and non-small cell lung cancer (NSCLC). Statins, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors used primarily for hyperlidpidemia, have been studied for their antiproliferative and pro-apoptotic effects. However, the effects of simvastatin on pemetrexed-induced apoptosis have not been investigated. In this study, we investigated whether combination treatment with pemetrexed and simvastatin potentiates the apoptotic activity above that is seen with either drug alone in malignant mesothelioma and NSCLC cells. We found that the combination of pemetrexed and simvastatin induced more extensive caspase-dependent apoptosis than either drug alone in malignant mesothelioma cells (MSTO-211) or NSCLC cells (A549). In addition, reactive oxygen species (ROS) generation in cells treated with both pemetrexed and simvastatin was markedly increased compared to cells treated with either pemetrexed or simvastatin alone. Combination treatment also increased the loss of mitochondrial membrane potential, increased cytosolic release of cytochrome c, and altered expression of inhibitor of apoptosis proteins (IAP) and B-cell lymphoma-2 (Bcl-2) families of apoptosis related proteins. On the other hand, pretreatment with N-acetylcysteine (NAC) prevented apoptosis and mitochondrial dysfunction by pemetrexed and simvastatin. In addition, Bim siRNA conferred protection against apoptosis induced by pemetrexed and simvastatin. These results suggest that combination of pemetrexed and simvastatin potentiates their apoptotic activity beyond that of either drug alone in malignant mesothelioma and lung cancer cells. This activity is mediated through ROS-dependent mitochondrial dysfunction and Bim induction.

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1	Jemal A, Siegel R, Xu J and Ward E: Cancer statistics, 2010. CA Cancer J Clin. 60:277–300. 2010. View Article : Google Scholar
2	Hotta K, Matsuo K, Ueoka H, Kiura K, Tabata M and Tanimoto M: Meta-analysis of randomized clinical trials comparing cisplatin to carboplatin in patients with advanced non-small-cell lung cancer. J Clin Oncol. 22:3852–3859. 2004. View Article : Google Scholar : PubMed/NCBI
3	Belli C, Fennell D, Giovannini M, Gaudino G and Mutti L: Malignant pleural mesothelioma: current treatments and emerging drugs. Expert Opin Emerg Drugs. 14:423–437. 2009. View Article : Google Scholar : PubMed/NCBI
4	Vogelzang NJ, Rusthoven JJ, Symanowski J, Denham C, Kaukel E, Ruffie P, Gatzemeier U, Boyer M, Emri S, Manegold C, Niyikiza C and Paoletti P: Phase III study of pemetrexed in combination with cisplatin versus cisplatin alone in patients with malignant pleural mesothelioma. J Clin Oncol. 21:2636–2644. 2003. View Article : Google Scholar : PubMed/NCBI
5	Scagliotti GV, Parikh P, von Pawel J, Biesma B, Vansteenkiste J, Manegold C, Serwatowski P, Gatzemeier U, Digumarti R, Zukin M, Lee JS, Mellemgaard A, Park K, Patil S, Rolski J, Goksel T, de Marinis F, Simms L, Sugarman KP and Gandara D: Phase III study comparing cisplatin plus gemcitabine with cisplatin plus pemetrexed in chemotherapy-naïve patients with advanced-stage non-small-cell lung cancer. J Clin Oncol. 26:3543–3551. 2008.PubMed/NCBI
6	Hong J, Kyung SY, Lee SP, Park JW, Jung SH, Lee JI, Park SH, Sym SJ, Park J, Cho EK, Shin DB and Lee JH: Pemetrexed versus gefitinib versus erlotinib in previously treated patients with non-small cell lung cancer. Korean J Intern Med. 25:294–300. 2010. View Article : Google Scholar : PubMed/NCBI
7	Shih C, Chen VJ, Gossett LS, Gates SB, MacKellar WC, Habeck LL, Shackelford KA, Mendelsohn LG, Soose DJ, Patel VF, Andis SL, Bewley JR, Rayl EA, Moroson BA, Beardsley GP, Kohler W, Ratnam M and Schultz RM: LY231514, a pyrrolo[2,3-d]pyrimidine-based antifolate that inhibits multiple folate-requiring enzymes. Cancer Res. 57:1116–1123. 1997.
8	Park CK, Kim KS, Oh IJ, Tseden-Ish M, Choi YD, Kwon YS, Kim YI, Lim SC and Kim YC: Efficacy of pemetrexed in relapsed non-small cell lung cancer and thymidylate synthase expression. Tuberc Respir Dis. 67:191–198. 2009. View Article : Google Scholar
9	Ramirez JM, Ocio EM, San Miguel JF and Pandiella A: Pemetrexed acts as an antimyeloma agent by provoking cell cycle blockade and apoptosis. Leukemia. 21:797–804. 2007.PubMed/NCBI
10	Lu X, Errington J, Curtin NJ, Lunec J and Newell DR: The impact of p53 status on cellular sensitivity to antifolate drugs. Clin Cancer Res. 7:2114–2123. 2001.PubMed/NCBI
11	Vandermeers F, Hubert P, Delvenne P, Mascaux C, Grigoriu B, Burny A, Scherpereel A and Willems L: Valproate, in combination with pemetrexed and cisplatin, provides additional efficacy to the treatment of malingnant mesothelioma. Clin Cancer Res. 15:2818–2828. 2009. View Article : Google Scholar : PubMed/NCBI
12	Goldstein JL and Brown MS: Regulation of the mevalonate pathway. Nature. 343:425–430. 1990. View Article : Google Scholar : PubMed/NCBI
13	Chan KK, Oza AM and Siu LL: The statins as anticancer agents. Clin Cancer Res. 9:10–19. 2003.
14	Demierre MF, Higgins PD, Gruber SB, Hawk E and Lippman SM: Statins and cancer prevention. Nat Rev Cancer. 5:930–942. 2005. View Article : Google Scholar : PubMed/NCBI
15	Holstein SA and Hohl RJ: Synergistic interaction of lovastatin and paclitaxel in human cancer cells. Mol Cancer Ther. 1:141–149. 2001.PubMed/NCBI
16	Feleszko W, Mlynarczuk I, Olszewska D, Jalili A, Grzela T, Lasek W, Hoser G, Korczak-Kowalska G and Jakobisiak M: Lovastatin potentiates antitumor activity of doxorubicin in murine melanoma via an apoptosis-dependent mechanism. Int J Cancer. 100:111–118. 2002. View Article : Google Scholar : PubMed/NCBI
17	Khanzada UK, Pardo OE, Meier C, Downward J, Seckl MJ and Arcaro A: Potent inhibition of small-cell lung cancer cell growth by simvastatin reveals selective functions of Ras isoforms in growth factor signaling. Oncogene. 25:877–887. 2006. View Article : Google Scholar : PubMed/NCBI
18	Kozar K, Kaminski R, Legat M, Kopec M, Nowis D, Skierski JS, Koronkiewicz M, Jakobisiak M and Golab J: Cerivastatin demonstrates enhanced antitumor activity against human breast cancer cell lines when used in combination with doxorubicin or cisplatin. Int J Oncol. 24:1149–1157. 2004.
19	Hwang KE, Park C, Kwon SJ, Kim YS, Park DS, Lee MK, Kim BR, Park SH, Yoon KH, Jeong ET and Kim HR: Synergistic induction of apoptosis by sulindac and simvastatin in A549 human lung cancer cells via reactive oxygen species-dependent mitochondrial dysfunction. Int J Oncol. 43:262–270. 2013.PubMed/NCBI
20	Sun JM, Lee KW, Kim JH, Kim YJ, Yoon HI, Lee JH, Lee CT and Lee JS: Efficacy and toxicity of pemetrexed as a third-line treatment of non-small cell lung cancer. Jpn J Clin Oncol. 39:27–32. 2009. View Article : Google Scholar : PubMed/NCBI
21	Kim KH, Song SY, Lim KH, Han SS, Kim SH, Cho JH, Park CW, Lee S and Lee HY: Interstitial pneumonitis after treatment with pemetrexed for non-small cell lung cancer. Cancer Res Treat. 45:74–77. 2013. View Article : Google Scholar : PubMed/NCBI
22	Wolf CM and Eastman A: The temporal relationship between protein phosphatase, mitochondrial cytochrome c release. Exp Cell Res. 247:505–513. 1999. View Article : Google Scholar : PubMed/NCBI
23	Elbashir SM, Harborth J, Lendeckel W, Yalcin A, Weber K and Tuschi T: Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature. 411:494–498. 2001. View Article : Google Scholar : PubMed/NCBI
24	Prevost GP, Pradines A, Brezak MC, Lonchampt MO, Viossat I, Ader I, Toulas C, Kasprzyk P, Gordon T, Favre G and Morgan B: Inhibition of human tumor cell growth in vivo by an orally bioavailable inhibitor of human farnesyltransferase, BIM-46228. Int J Cancer. 91:718–722. 2001. View Article : Google Scholar : PubMed/NCBI
25	Song JY, Kim CS, Lee JH, Jang SJ, Lee SW, Hwang JJ, Lim C, Lee G, Seo J, Cho SY and Choi J: Dual inhibition of MEK 1/2 and EGFR synergistically induces caspase-3-dependent apoptosis in EGFR inhibitor-resistant lung cancer cells via BIM upregulation. Invest New Drugs. 31:1458–1465. 2013. View Article : Google Scholar : PubMed/NCBI
26	Schulze-Bergkamen H and Krammer PH: Apotosis in cancer-implications for therapy. Semin Oncol. 31:90–119. 2004. View Article : Google Scholar
27	Copeland WC, Wachsman JT, Johnson FM and Penta JS: Mitochondrial DNA alterations in cancer. Cancer Invest. 20:557–569. 2002. View Article : Google Scholar : PubMed/NCBI
28	Kim HR, Yang SH and Jeong ET: Combination treatment with arsenic trioxide and sulindac induces apoptosis of NCI-H157 human lung carcinoma cells via ROS generation with mitochondrial dysfunction. Tuberc Respir Dis. 59:30–38. 2005.
29	Frank S, Gaume B, Bergmann-Leitner ES, Leitner WW, Robert EG, Catez F, Smith CL and Youle RJ: The role of dynamic-related protein 1, a mediator of mitochondrial fission, in apoptosis. Dev Cell. 1:515–525. 2001. View Article : Google Scholar : PubMed/NCBI
30	Karbowski M, Lee YJ, Gaume B, Jeong SY, Frank S, Nechushtan A, Santel A, Fuller M, Smith CL and Youle RJ: Spatial and temporal association of Bax with mitochondrial fission sites, Drp1, and Mfn2 during apoptosis. J Cell Biol. 159:931–938. 2002. View Article : Google Scholar : PubMed/NCBI
31	Green DR and Reed JC: Mitochondria and apoptosis. Science. 281:1309–1312. 1998. View Article : Google Scholar
32	Li LY, Luo X and Wang X: Endonuclease G is an apoptotic DNase when released from mitochondria. Nature. 412:95–99. 2001. View Article : Google Scholar : PubMed/NCBI
33	Tournier C, Hess P, Yang DD, Xu J, Turner TK, Nimnual A, Bar-Sagi D, Jones SN, Flavell RA and Davis RJ: Requirement of JNK for stress-induced activation of the cytochrome c-mediated death pathway. Science. 288:870–874. 2000. View Article : Google Scholar : PubMed/NCBI
34	Burdon RH: Control of cell proliferation by reactive oxygen species. Biochem Soc Trans. 24:1028–1032. 1996.PubMed/NCBI
35	Buque A, Muhialdin JSh, Munoz A, Calvo B, Carrera S, Aresti U, Sancho A, Rubio I and Lopez-Vivinco G: Molecular mechanism implicated in pemetrexed-induced apoptosis in human melanoma cells. Mol Cancer. 11:252012. View Article : Google Scholar : PubMed/NCBI
36	Fiers W, Beyaert R, Declercq W and Vandenabeele P: More than one way to die: apoptosis, necrosis and reactive oxygen damage. Oncogene. 18:7719–7730. 1999. View Article : Google Scholar