Planta Med 2013; 79(09): 775-781
DOI: 10.1055/s-0032-1328554
Biological and Pharmacological Activity
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Arsenic Trioxide-Enhanced, Matrine-Induced Apoptosis in Multiple Myeloma Cell Lines

Qinghong Yu
1   Department of Hematology, The First Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou, ZheJiang, China
,
Binhai Chen
3   Department of Oncology, The Second Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou, ZheJiang, China
,
Xiang Zhang
1   Department of Hematology, The First Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou, ZheJiang, China
,
WenBin Qian
2   Department of Hematology, Institute of Hematology, The First Affiliated Hospital, ZheJiang University College of Medicine, Hangzhou, ZheJiang, China
,
Baodong Ye
1   Department of Hematology, The First Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou, ZheJiang, China
,
Yuhong Zhou
1   Department of Hematology, The First Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou, ZheJiang, China
› Author Affiliations
Further Information

Publication History

received 26 December 2012
revised 20 March 2013

accepted 09 April 2013

Publication Date:
22 May 2013 (online)

Abstract

Matrine and arsenic trioxide are monomers used in traditional Chinese medicine possessing anti-myeloma activities. In this study, we evaluated the effects and mechanisms of matrine, arsenic trioxide, and their combination therapy on the proliferation and apoptosis of the myeloma cell lines RPMI8226 and U266. The effects of growth inhibition were measured by MTT, and apoptotic cells were analyzed by Hoechst 33258 staining and flow cytometry. The levels of caspase-3, poly (ADP-ribose) polymerase (a DNA repair enzyme), Bcl-2 and survivin (antiapoptotic signaling proteins), Bim (a proapoptotic signaling protein), total AKT, and phosphorylated AKT were evaluated by Western blot. Matrine significantly inhibited proliferation of RPMI8226 and U266 cell lines in a dose- and time-dependent manner with an IC50 at 24 h of 2.25 g/L and 2.18 g/L, and at 48 h of 1.64 g/L and 1.58 g/L, respectively. Arsenic trioxide also displayed a dose- and time-dependent inhibition of growth of multiple myeloma cell lines, and synergistic effects occurred when the two were combined. Matrine (0.5, 1.0 g/L) and arsenic trioxide (2, 4 ug/mL) induced the apoptosis of myeloma cells; more early-stage apoptotic cells were seen with the combination therapy (matrine 0.5 g/L plus arsenic trioxide 2 ug/mL and matrine 1.0 g/L plus arsenic trioxide 4 ug/mL). Activation of caspase-3 and poly (ADP-ribose) polymerase, upregulation of Bim expression, downregulation of Bcl-2, survivin expression, as well as inhibition of phosphorylated AKT related to matrine (0, 0.25, 0.5, 1.0, and 2.0 g/L)-mediated apoptosis, and the effects were enhanced when arsenic trioxide (8 ug/mL) was combined with matrine (1.0 g/L). In conclusion, matrine displayed anti-myeloma effects through apoptotic induction, and arsenic trioxide had synergistic effects with matrine enhancing matrine-induced apoptosis.

Supporting Information

 
  • References

  • 1 Zhang LP, Jiang JK, Tam JW, Zhang Y, Liu XS, Xu XR, Liu BZ, He YJ. Effects of matrine on proliferation and differentiation in K562 cells. Leuk Res 2001; 25: 793-800
  • 2 Han Y, Zhang S, Wu J, Yu K, Zhang Y, Yin L, Bi L. Matrine induces apoptosis of human multiple myeloma cells via activation of the mitochondrial pathway. Leuk Lymphoma 2010; 517: 1337-1346
  • 3 Wu JB, Zhang SH, Han YX, Xiong SD, Ye AF, Tan YX. Effects of matrine on the apoptosis and expression of adhesion molecue in multiple myeloma RMPI8226 cells. Zhongguo Shi Yan Xue Ye Xue Za Zhi 2008; 16: 93-96
  • 4 Sun M, Cao H, Sun L, Dong S, Bian Y, Han J, Zhang L, Ren S, Hu Y, Liu C, Xu L, Liu P. Antitumor activities of kushen: literature review. Evid Based Complement Alternat Med 2012; 2012: 373219
  • 5 Zhang W, Yang X. Clinical study of compound kuh-seng injection combined with chemotherapy for multiple myeloma. J TCM Univ Hunan 2010; 30: 68-75
  • 6 Guo Y. Observation on treating acute leukemia adjuvant chemotherapy in compound matrine injection. Clin J Chin Med 2011; 3: 98-100
  • 7 Zheng Y, Zheng T. Efficacy observation of compound kushen injection combined with chemotherapy in the treatment of non-hodgkinʼs lymphoma. Pharm Clin Chin Mater Med 2011; 2: 55-56
  • 8 Chen J, Yu Y, Wang X, Zhou Y, Bao F, Wu X. Clinical observation of adjuvant therapy of compound Sophora flavescens injection for multiple myeloma bone pain. Chin Pharm 2011; 22: 1880-1882
  • 9 Zhang Z. Effects of matrine injection in treating 44 cases of primary carcinoma of the liver. Guangxi Tradit Med 1998; 21: 19-20
  • 10 Zhou Y. Clinical observation on the efficacy of mateling injection combined with abdominal chemotherapy in treating malignant ascites. J Emerg Tradit Chin Med 2004; 13: 286-287
  • 11 Baysan A, Yel L, Gollapudi S, Su H, Gupta S. Arsenic trioxide induces apoptosis via the mitochondrial pathway by upregulating the expression of Bcl-2 and Bim in human B cells. Int J Oncol 2007; 30: 313-318
  • 12 Kircelli F, Akay C, Gazitt Y. Arsenic trioxide induces p 53-dependent apoptotic signals in myeloma cells with SiRNA-silenced p 53: MAP kinase pathway is preferentially activated in cells expressing inactivated p 53. Int J Oncol 2007; 30: 993-1001
  • 13 Liu Q, Hilsenbeck S, Gazitt Y. Arsenic trioxide-induced apoptosis in myeloma cells: p 53-dependent G1 or G2/M cell cycle arrest, activation of caspase-8 or caspase-9, and synergy with APO2/TRAIL. Blood 2003; 101: 4078-4087
  • 14 Wu X, Shi J, Wu Y, Tao Y, Hou J, Meng X, Hu X, Han Y, Jiang W, Tang S, Zangari M, Tricot G, Zhan F. Arsenic trioxide-mediated growth inhibition of myeloma cells is associated with an extrinsic or intrinsic signaling pathway through activation of TRAIL or TRAIL receptor 2. Cancer Biol Ther 2010; 10: 1201-1214
  • 15 Yu QH, Zhan R, Huang HB. Mechanisms underlying the effect of arsenic trioxide on proliferation inhibition and apoptosis induction in myeloma cell line u266. Zongguo Shi Yan Xue Ye Xue Za Zhi 2007; 15: 982-985
  • 16 Kajiguchi T, Yamamoto K, Iida S, Ueda R, Emi N, Naoe T. Sustained activation of c-jun-N-terminal kinase plays a critical role in arsenic trioxide-induced cell apoptosis in multiple myeloma cell lines. Cancer Sci 2006; 97: 540-545
  • 17 Morales AA, Gutman D, Lee KP, Boise LH. BH3-only proteins Noxa, Bmf, and Bim are necessary for arsenic trioxide-induced cell death in myeloma. Blood 2008; 111: 5152-5162
  • 18 Morales AA, Gutman D, Cejas PJ, Lee KP, Boise LH. Reactive oxygen species are not required for an arsenic trioxide-induced antioxidant response or apoptosis. J Biol Chem 2009; 284: 12886-12895
  • 19 Park WH, Seol JG, Kim ES, Hyun JM, Jung CW, Lee CC, Kim BK, Lee YY. Arsenic trioxide-mediated growth inhibition in MC/CAR myeloma cells via cell cycle arrest in association with induction of cyclin-dependent kinase inhibitor, p 21, and apoptosis. Cancer Res 2000; 60: 3065-3071
  • 20 Wang MM, Dou HJ, Zou LF, Zhu Q, Ren ZH, Hu JP. Correlation of cell cycle alteration to SOCS-1 gene demethylation induced by arsenic trioxide in myeloma cell lines. Ai Zheng 2008; 27: 1150-1154
  • 21 Fu HY, Shen JZ. Hypermethylation of CpG island of p16 gene and arsenic trioxide induced p 16 gene demethylation in multiple myeloma. Zhonghua Nei Ke Za Zhi 2005; 44: 411-414
  • 22 Fu HY, Sheng JZ, Sheng SF, Zhou HR. n-MSP detection of p16 gene demethylation and transcription in human multiple myeloma U266 cell line induced by arsenic trioxide. Zhongguo Shi Yan Xue Ye Xue Za Zhi 2007; 15: 79-85
  • 23 Qu X, Du J, Zhang C, Fu W, Xi H, Zou J, Hou J. Arsenic trioxide exerts antimyeloma effects by inhibiting activity in the cytoplasmic substrates of histone deacetylase 6. PLoS One 2012; 7: e32215
  • 24 Deaglio S, Canella D, Baj G, Arnulfo A, Waxman S, Malavasi F. Evidence of an immunologic mechanism behind the therapeutical effects of arsenic trioxide (As(2)O(3)) on myeloma cells. Leuk Res 2001; 25: 227-235
  • 25 Hussein MA. Arsenic trioxide: a new immunomodulatory agent in the management of multiple myeloma. Med Oncol 2001; 18: 239-242
  • 26 Wang Y, Hu Y, Sun C, Zhang X, He W. Mechanism of arsenic trioxide inhibiting angiogenesis in multiple myeloma. J Huazhong Univ Sci Technolog Med Sci 2006; 26: 43-46
  • 27 Hofmeister CC, Jansak B, Denlinger N, Kraut EH, Benson DM, Farag SS. Phase II clinical trial of arsenic trioxide with liposomal doxorubicin, vincristine, and dexamethasone in newly diagnosed multiple myeloma. Leuk Res 2008; 32: 1295-1298
  • 28 Sanaat Z, Rezazadeh M, Gharamaleki JV, Ziae JE, Esfahani A. Arsenic trioxide in patients with refractory multiple myeloma: a prospective, phase II, single-arm study. Acta Med Iran 2011; 49: 504-508
  • 29 Berenson JR, Matous J, Swift RA, Mapes R, Morrison B, Yeh HS. A phase I/II study of arsenic trioxide/bortezomib/ascorbic acid combination therapy for the treatment of relapsed or refractory multiple myeloma. Clin Cancer Res 2007; 13: 1762-1768
  • 30 Abou-Jawde RM, Reed J, Kelly M, Walker E, Andresen S, Baz R, Karam MA, Hussein M. Efficacy and safety results with the combination therapy of arsenic trioxide, dexamethasone, and ascorbic acid in multiple myeloma patients: a phase 2 trial. Med Oncol 2006; 23: 263-272
  • 31 Sharma M, Khan H, Thall PF, Orlowski RZ, Bassett jr. RL, Shah N, Bashir Q, Parmar S, Wang M, Shah JJ, Hosing CM, Popat UR, Giralt SA, Champlin RE, Qazilbash MH. A randomized phase 2 trial of a preparative regimen of bortezomib, high-dose melphalan, arsenic trioxide, and ascorbic acid. Cancer 2012; 118: 2507-2515
  • 32 Liu X, Jiang J. Molecular mechanism of matrine induced apoptosis in leukemia K562 cells. Am J Chin Med 2006; 34: 1095-1103
  • 33 Chui CH, Lau FY, Tang JC, Kan KL, Cheng GY, Wong RS, Kok SH, Lai PB, Ho R, Gambari R, Chan AS. Activities of fresh juice of Scutellaria barbata and warmed water extract of Radix Sophorae tonkinensis on anti-proliferation and apoptosis of human cancer cell lines. Int J Mol Med 2005; 16: 337-341
  • 34 Yu P, Liu Q, Liu K, Yagasaki K, Wu E, Zhang G. Matrine suppresses breast cancer cell proliferation and invasion via VEGF-Akt-NF-kappaB signaling. Cytotechnology 2009; 59: 219-229
  • 35 Zhang S, Zhang Y, Zhuang Y, Wang J, Ye J, Zhang S, Wu J, Yu K, Han Y. Matrine induces apoptosis in human acute myeloid leukemia cells via the mitochondrial pathway and akt inactivation. PLoS One 2012; 7: e46853
  • 36 Campbell RA, Sanchez E, Steinberg JA, Baritaki S, Gordon M, Wang C, Shalitin D, Chen H, Pang S, Bonavida B, Said J, Berenson JR. Antimyeloma effects of arsenic trioxide are enhanced by melphalan, bortezomib and ascorbic acid. Br J Haematol 2007; 138: 467-478
  • 37 Wen J, Feng Y, Huang W, Chen H, Liao B, Rice L, Preti HA, Kamble RT, Zu Y, Ballon DJ, Chang CC. Enhanced antimyeloma cytotoxicity by the combination of arsenic trioxide and bortezomib is further potentiated by p 38 MAPK inhibition. Leuk Res 2010; 34: 85-92
  • 38 Zhou L, Hou J, Fu W, Wang D, Yuan Z, Jiang H. Arsenic trioxide and 2-methoxyestradiol reduce beta-catenin accumulation after proteasome inhibition and enhance the sensitivity of myeloma cells to Bortezomib. Leuk Res 2008; 32: 1674-1683
  • 39 Ge QF, Ouyang GF, Chen Y, Zhang Y, Mu QT, Lu Y. Effects of arsenic trioxide combined with bortezomib on apoptosis of multiple myeloma cell line KM3 and its mechanisms. Zhongguo Shi Yan Xue Ye Xue Za Zhi 2012; 20: 112-115
  • 40 Wen J, Cheng HY, Feng Y, Rice L, Liu S, Mo A, Huang J, Zu Y, Ballon DJ, Chang CC. P38 MAPK inhibition enhancing ATO-induced cytotoxicity against multiple myeloma cells. Br J Haematol 2008; 140: 169-180
  • 41 Romagnoli M, Séveno C, Wuillème-Toumi S, Amiot M, Bataille R, Minvielle S, Barillé-Nion S. The imbalance between survivin and Bim mediates tumour growth and correlates with poor survival in patients with multiple myeloma. Br J Haematol 2009; 145: 180-189
  • 42 Tsubaki M, Satou T, Itoh T, Imano M, Komai M, Nishinobo M, Yamashita M, Yanae M, Yamazoe Y, Nishida S. Overexpression of MDR1 and survivin, and decreased Bim expression mediate multidrug-resistance in multiple myeloma cells. Leuk Res 2012; 36: 1315-1322
  • 43 Hsu J, Shi Y, Krajewski S, Renner S, Fisher M, Reed JC, Franke TF, Lichtenstein A. The AKT kinase is activated in multiple myeloma tumor cells. Blood 2001; 98: 2853-2855
  • 44 Tu Y, Gardner A, Lichtenstein A. The phosphatidylinositol 3-kinase/AKT kinase pathway in multiple myeloma plasma cells: roles in cytokine-dependent survival and proliferative responses. Cancer Res 2000; 60: 6763-6770
  • 45 Meng H, Yang C, Jin J, Zhou Y, Qian W. Homoharringtonine inhibits the AKT pathway and induces in vitro and in vivo cytotoxicity in human multiple myeloma cells. Leuk Lymphoma 2008; 49: 1954-1962