Effects of gambogic acid on the activation of caspase-3 and downregulation of SIRT1 in RPMI-8226 multiple myeloma cells via the accumulation of ROS

Yang,Li-Jing; Chen,Yan; He,Jing; Yi,Sha; Wen,Lu; Zhao,Shuai; Cui,Guo-Hui

doi:10.3892/ol.2012.634

Effects of gambogic acid on the activation of caspase-3 and downregulation of SIRT1 in RPMI-8226 multiple myeloma cells via the accumulation of ROS

Authors:
- Li-Jing Yang
- Yan Chen
- Jing He
- Sha Yi
- Lu Wen
- Shuai Zhao
- Guo-Hui Cui
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Affiliations: Department of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, P.R. China
Published online on: March 6, 2012 https://doi.org/10.3892/ol.2012.634
Pages: 1159-1165

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Abstract

Multiple myeloma (MM) is the second most commonly diagnosed hematologic malignancy. Although new drugs, including bortezomib and lenalidomide, have improved the treatment landscape for MM patients, MM remains incurable. Therefore, screening for novel anti-myeloma drugs is necessary. Gambogic acid (GA), the main active ingredient of gamboges secreted from the Garcinia hanburryi tree, has been reported to exhibit potent anticancer activity in certain solid tumors and hematological malignancies, while there are few studies that are available concerning its effects on MM cells. In the present study, we investigated the anticancer activity of GA on the MM RPMI-8226 cells and further studied the underlying mechanisms by which GA affected the cells. RPMI-8226 cells were cultured and the effect of GA on cell proliferation was analyzed using MTT assay. Hoechst 33258 staining was used to visualize nuclear fragmentation, and reactive oxygen species (ROS) levels were detected. GA was found to have a significant, dose-dependent effect on growth inhibition and apoptosis induction in RPMI-8226 cells. This activity is associated with the accumulation of ROS, which contributes to the activation of caspase-3 and the cleavage of poly (ADP-ribose) polymerase (PARP), accompanied with apoptosis in RPMI-8226 cells treated with GA. Mammalian SIRT1, as the closest homolog of the yeast Sir2, was extensively involved in regulating cell processes, including cell senescence, aging and neuronal protection, as well as having anti-apoptotic properties. Moreover, SIRT1 overexpression has been shown to protect cancer cells from chemotherapy and ionizing radiation. In the present study, we demonstrated that GA has the potential to downregulate the expression of SIRT1 via ROS accumulation. In conclusion, our study found that GA is able to induce apoptosis in RPMI‑8226 cells via ROS accumulation followed by caspase-3 activation, PARP cleavage and SIRT1 downregulation. These results suggest that GA may have the potential to not only induce apoptosis in MM cells, but also to decrease the relapse rate of MM.

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1	Kasenda B, Ruckert A, Farthmann J, et al: Management of multiple myeloma in pregnancy: strategies for a rare challenge. Clin Lymphoma Myeloma Leuk. 11:190–197. 2011. View Article : Google Scholar : PubMed/NCBI
2	Siegel DS, Vij R and Jakubowiak AJ: Clinical roundtable monograph. Emerging treatment options for relapsed and refractory multiple myeloma. Clin Adv Hematol Oncol. 9:1–15. 2011.PubMed/NCBI
3	Gu H, Rao S, Zhao J, et al: Gambogic acid reduced bcl-2 expression via p53 in human breast MCF-7 cancer cells. J Cancer Res Clin Oncol. 135:1777–1782. 2009. View Article : Google Scholar : PubMed/NCBI
4	Xu X, Liu Y, Wang L, et al: Gambogic acid induces apoptosis by regulating the expression of Bax and Bcl-2 and enhancing caspase-3 activity in human malignant melanoma A375 cells. Int J Dermatol. 48:186–192. 2009. View Article : Google Scholar : PubMed/NCBI
5	Cui G, Shu W, Wu Q and Chen Y: Effect of Gambogic acid on the regulation of hERG channel in K562 cells in vitro. J Huazhong Univ Sci Technolog Med Sci. 29:540–545. 2009. View Article : Google Scholar : PubMed/NCBI
6	Wang Y, Chen Y, Chen Z, Wu Q, Ke WJ and Wu QL: Gambogic acid induces death inducer-obliterator 1-mediated apoptosis in Jurkat T cells. Acta Pharmacol Sin. 29:349–354. 2008. View Article : Google Scholar : PubMed/NCBI
7	Rong JJ, Hu R, Song XM, et al: Gambogic acid triggers DNA damage signaling that induces p53/p21 (Waf1/CIP1) activation through the ATR-Chk1 pathway. Cancer Lett. 296:55–64. 2010. View Article : Google Scholar : PubMed/NCBI
8	Nanduri J, Wang N, Bergson P, Yuan G, Ficker E and Prabhakar NR: Mitochondrial reactive oxygen species mediate hypoxic down-regulation of hERG channel protein. Biochem Biophys Res Commun. 373:309–314. 2008. View Article : Google Scholar : PubMed/NCBI
9	Liu B, Chen Y and St Clair DK: ROS and p53: a versatile partnership. Free Radic Biol Med. 44:1529–1535. 2008. View Article : Google Scholar : PubMed/NCBI
10	Azad N, Iyer A, Vallyathan V, et al: Role of oxidative/nitrosative stress-mediated Bcl-2 regulation in apoptosis and malignant transformation. Ann NY Acad Sci. 1203:1–6. 2010. View Article : Google Scholar : PubMed/NCBI
11	Borutaite V and Brown GC: Caspases are reversibly inactivated by hydrogen peroxide. FEBS Lett. 500:114–118. 2001. View Article : Google Scholar : PubMed/NCBI
12	Chen YC, Shen SC and Tsai SH: Prostaglandin D(2) and J(2) induce apoptosis in human leukemia cells via activation of the caspase 3 cascade and production of reactive oxygen species. Biochim Biophys Acta. 1743:291–304. 2005. View Article : Google Scholar : PubMed/NCBI
13	Nie F, Zhang X, Qi Q, et al: Reactive oxygen species accumulation contributes to gambogic acid-induced apoptosis in human hepatoma SMMC-7721 cells. Toxicology. 260:60–67. 2009. View Article : Google Scholar : PubMed/NCBI
14	Scherz-Shouval R and Elazar Z: Regulation of autophagy by ROS: physiology and pathology. Trends Biochem Sci. 36:30–38. 2011. View Article : Google Scholar : PubMed/NCBI
15	Bokoch GM: Regulation of the human neutrophil NADPH oxidase by the Rac GTP-binding proteins. Curr Opin Cell Biol. 6:212–218. 1994. View Article : Google Scholar : PubMed/NCBI
16	Sundaresan M, Yu ZX, Ferrans VJ, et al: Regulation of reactive-oxygen-species generation in fibroblasts by Rac1. Biochem J. 318(Pt 2): 379–382. 1996.PubMed/NCBI
17	Terada LS: Specificity in reactive oxidant signaling: think globally, act locally. J Cell Biol. 174:615–623. 2006. View Article : Google Scholar : PubMed/NCBI
18	Jones DP: Redefining oxidative stress. Antioxid Redox Signal. 8:1865–1879. 2006. View Article : Google Scholar
19	Valko M, Leibfritz D, Moncol J, Cronin MT, Mazur M and Telser J: Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol. 39:44–84. 2007. View Article : Google Scholar : PubMed/NCBI
20	Chen Y, Jungsuwadee P, Vore M, Butterfield DA and St Clair DK: Collateral damage in cancer chemotherapy: oxidative stress in nontargeted tissues. Mol Interv. 7:147–156. 2007. View Article : Google Scholar : PubMed/NCBI
21	Smith BC, Hallows WC and Denu JM: Mechanisms and molecular probes of sirtuins. Chem Biol. 15:1002–1013. 2008. View Article : Google Scholar : PubMed/NCBI
22	Luo J, Nikolaev AY, Imai S, et al: Negative control of p53 by Sir2alpha promotes cell survival under stress. Cell. 107:137–148. 2001. View Article : Google Scholar : PubMed/NCBI
23	Zeng R, He J, Peng J, et al: The time-dependent autophagy protects against apoptosis with possible involvement of Sirt1 protein in multiple myeloma under nutrient depletion. Ann Hematol. 91:407–417. 2012. View Article : Google Scholar : PubMed/NCBI
24	Olmos Y, Brosens JJ and Lam EW: Interplay between SIRT proteins and tumour suppressor transcription factors in chemotherapeutic resistance of cancer. Drug Resist Updat. 14:35–44. 2011. View Article : Google Scholar : PubMed/NCBI
25	Chauhan D, Bandi M, Singh AV, et al: Preclinical evaluation of a novel SIRT1 modulator SRT1720 in multiple myeloma cells. Br J Haematol. 155:588–598. 2011. View Article : Google Scholar : PubMed/NCBI
26	Lazebnik YA, Kaufmann SH, Desnoyers S, Poirier GG and Earnshaw WC: Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature. 371:346–347. 1994. View Article : Google Scholar : PubMed/NCBI
27	Prasad S, Pandey MK, Yadav VR and Aggarwal BB: Gambogic acid inhibits STAT3 phosphorylation through activation of protein tyrosine phosphatase SHP-1: potential role in proliferation and apoptosis. Cancer Prev Res (Phila). 4:1084–1094. 2011. View Article : Google Scholar : PubMed/NCBI
28	Degterev A, Boyce M and Yuan J: A decade of caspases. Oncogene. 22:8543–8567. 2003. View Article : Google Scholar : PubMed/NCBI
29	Salvesen GS and Abrams JM: Caspase activation stepping on the gas or releasing the brakes? Lessons from humans and flies. Oncogene. 23:2774–2784. 2004. View Article : Google Scholar : PubMed/NCBI
30	Circu ML and Aw TY: Reactive oxygen species, cellular redox systems, and apoptosis. Free Radic Biol Med. 48:749–762. 2010. View Article : Google Scholar : PubMed/NCBI
31	Slee EA, Harte MT, Kluck RM, et al: Ordering the cytochrome c-initiated caspase cascade: hierarchical activation of caspases-2, -3, -6, -7, -8, and -10 in a caspase-9-dependent manner. J Cell Biol. 144:281–292. 1999. View Article : Google Scholar : PubMed/NCBI
32	Fraga MF, Agrelo R and Esteller M: Cross-talk between aging and cancer: the epigenetic language. Ann NY Acad Sci. 1100:60–74. 2007. View Article : Google Scholar : PubMed/NCBI
33	Fraga MF and Esteller M: Epigenetics and aging: the targets and the marks. Trends Genet. 23:413–418. 2007. View Article : Google Scholar : PubMed/NCBI
34	Hida Y, Kubo Y, Murao K and Arase S: Strong expression of a longevity-related protein, SIRT1, in Bowen's disease. Arch Dermatol Res. 299:103–106. 2007. View Article : Google Scholar : PubMed/NCBI
35	Lim CS: Human SIRT1: a potential biomarker for tumorigenesis? Cell Biol Int. 31:636–637. 2007. View Article : Google Scholar : PubMed/NCBI
36	Chu F, Chou PM, Zheng X, Mirkin BL and Rebbaa A: Control of multidrug resistance gene mdr1 and cancer resistance to chemotherapy by the longevity gene sirt1. Cancer Res. 65:10183–10187. 2005. View Article : Google Scholar : PubMed/NCBI
37	Matsushita N, Takami Y, Kimura M, et al: Role of NAD-dependent deacetylases SIRT1 and SIRT2 in radiation and cisplatin-induced cell death in vertebrate cells. Genes Cells. 10:321–332. 2005. View Article : Google Scholar : PubMed/NCBI
38	Brunet A, Sweeney LB, Sturgill JF, et al: Stress-dependent regulation of FOXO transcription factors by the SIRT1 deacetylase. Science. 303:2011–2015. 2004. View Article : Google Scholar : PubMed/NCBI
39	Zhao G, Cui J, Zhang JG, et al: SIRT1 RNAi knockdown induces apoptosis and senescence, inhibits invasion and enhances chemosensitivity in pancreatic cancer cells. Gene Ther. 18:920–928. 2011. View Article : Google Scholar : PubMed/NCBI