Abstract
Purpose
In a recent publication, we have shown that dihydroartemisinin (DHA), a derivative of antimalaria drug artemisinin, inhibits growth of pancreatic cancer cells in vitro and in vivo mediated by its anti-proliferative and pro-apoptotic effects. As it has been shown that the apoptosis might be induced due to cell cycle arrest, and that transcriptional factor nuclear factor-kappa B (NF-κB) plays vital roles in the apoptosis of pancreatic cancer cells, we extend our study to investigate the effects of DHA on cell cycle progression and NF-κB activity in pancreatic cancer cells to further reveal the anticancer effects of DHA on pancreatic cancer.
Methods
Cell cycle progression was determined by propidium iodide staining and flow cytometry. Changes in the expression of cell cycle-associated proteins were detected using Western blot analysis. Measurement of NF-κB activity was performed with immunoblot analyzing the nuclear protein expression of NF-κB/p65 and ELISA detecting the NF-κB DNA-binding activity.
Results
The treatment with DHA resulted in a dose-dependent G0/G1 cell cycle arrest and regulated the expression of some cyclins, cdks and cdk inhibitors that involved in the G0/G1 cell cycle progression such as cyclin E, cdk2, cdk4 and p27Kip1 in pancreatic cancer BxPC-3 and AsPC-1 cells. The translocation and DNA-binding activity of NF-κB were inhibited in DHA-treated cells in a dose-dependent manner, indicated the inactivation effects of DHA in pancreatic cancer cells.
Conclusions
Together with our previous observations, our data show that DHA induces cell cycle arrest and apoptosis in pancreatic cancer cells, and this effect might be due to inhibition of NF-κB signaling. We suggest that DHA could be developed as a novel agent against pancreatic cancer.
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References
Aggarwal BB (2004) Nuclear factor-κB: the enemy within. Cancer Cell 6:203–208. doi:10.1016/j.ccr.2004.09.003
Aldieri E, Atragene D, Bergandi L, Riganti C, Costamagna C, Bosia A, Ghigo D (2003) Artemisinin inhibits inducible nitric oxide synthase and nuclear factor NF-kB activation. FEBS Lett 552(2–3):141–144. doi:10.1016/S0014-5793(03)00905-0
Chen HH, Zhou HJ, Fang X (2003) Inhibition of human cancer cell line growth and human umbilical vein endothelial cell angiogenesis by artemisinin derivatives in vitro. Pharmacol Res 48:231–236. doi:10.1016/S1043-6618(03)00107-5
Chen H, Sun B, Pan S, Jiang H, Sun X (2009) Dihydroartemisinin inhibits growth of pancreatic cancer cells in vitro and in vivo. Anticancer Drugs 20(2):131–140. doi:10.1097/CAD.0b013e3283212ade
Dai Y, Grant S (2004) Small molecule inhibitors targeting cyclin-dependent kinases as anticancer agents. Curr Oncol Rep 6:123–130. doi:10.1007/s11912-004-0024-3
Derradji H, Baatout S (2003) Apoptosis: a mechanism of cell suicide. In Vivo 17:185–192
Devault A, Cavadore JC, Fesquet D, Labbé JC, Lorca T, Picard A, Strausfeld U, Dorée M (1991) Concerted roles of cyclin A, cdc25+ mitotic inducer, and type 2A phosphatase in activating the cyclin B/cdc2 protein kinase at the G2/M phase transition. Cold Spring Harb Symp Quant Biol 56:503–513. doi:10.1101/SQB.1991.056.01.057
Efferth T, Marschall M, Wang X, Huong SM, Hauber I, Olbrich A, Kronschnabl M, Stamminger T, Huang ES (2002) Antiviral activity of artesunate towards wild-type, recombinant, and ganciclovir-resistant human cytomegaloviruses. J Mol Med 80(4):233–242. doi:10.1007/s00109-001-0300-8
Efferth T, Sauerbrey A, Olbrich A, Gebhart E, Rauch P, Weber HO, Hengstler JG, Halatsch ME, Volm M, Tew KD, Ross DD, Funk JO (2003) Molecular modes of action of artesunate in tumor cell lines. Mol Pharmacol 64(2):382–394. doi:0026-895X/03/6402-382-394
Fujioka S, Sclabas GM, Schmidt C, Frederick WA, Dong QG, Abbruzzese JL, Evans DB, Baker C, Chiao PJ (2003) Function of nuclear factor κB in pancreatic cancer metastasis. Clin Cancer Res 9:346–354
Fujita T, Felix K, Pinkaew D, Hutadilok-Towatanab N, Liu Z, Fujise K (2008) Human fortilin is a molecular target of dihydroartemisinin. FEBS Lett 582:1055–1060. doi:10.1016/j.febslet.2008.02.055
Greten FR, Weber CK, Greten TF, Schneider G, Wagner M, Adler G, Schmid RM (2002) Stat3 and NF-κB activation prevents apoptosis in pancreatic carcinogenesis. Gastroenterology 123:2052–2063. doi:10.1053/gast.2002.37075
Hartwell LH, Kastan MB (1994) Cell cycle control and cancer. Science 266:1821–1828. doi:10.1126/science.7997877
Hou J, Wang D, Zhang R, Wang H (2008) Experimental therapy of hepatoma with artemisinin and its derivatives: in vitro and in vivo activity, chemosensitization, and mechanisms of action. Clin Cancer Res 14:5519–5530. doi:10.1158/1078-0432.CCR-08-0197
Huang XJ, Ma ZQ, Zhang WP, Lu YB, Wei EQ (2007) Dihydroartemisinin exerts cytotoxic effects and inhibits hypoxia inducible factor-1alpha activation in C6 glioma cells. J Pharm Pharmacol 59:849–856. doi:10.1211/jpp.59.6.0011
Jemal A, Murray T, Ward E, Samuels A, Tiwari RC, Ghafoor A, Feuer EJ, Thun MH (2005) Cancer statistics, 2005. CA Cancer J Clin 55(1):10–30. doi:10.3322/canjclin.55.1.10
Jiao Y, Ge CM, Meng QH, Cao JP, Tong J, Fan SJ (2007) Dihydroartemisinin is an inhibitor of ovarian cancer cell growth. Acta Pharmacol Sin 28:1045–1056. doi:10.1111/j.1745-7254.2007.00612.x
Karin M (2006) Nuclear factor-κB in cancer development and progression. Nature 441:431–436. doi:10.1038/nature04870
Karin M, Greten FR (2005) NF-κB: linking inflammation and immunity to cancer development and progression. Nat Rev Immunol 5:749–759. doi:10.1038/nri1703
Kim SJ, Kim MS, Lee JW, Lee CH, Yoo H, Shin SH, Park MJ, Lee SH (2006) Dihydroartemisinin enhances radiosensitivity of human glioma cells in vitro. J Cancer Res Clin Oncol 132:129–135. doi:10.1007/s00432-005-0052-x
Kruidering M, Evan GI (2000) Caspase-8 in apoptosis: the beginning of “the end”? IUBMB Life 50:85–90. doi:10.1080/713803693
Li D, Xie K, Wolff R, Abbruzzese JL (2004) Pancreatic cancer. Lancet 363:1049–1057. doi:10.1016/S0140-6736(04)15841-8
Liptay S, Weber CK, Ludwig L, Wagner M, Adler G, Schmid RM (2003) Mitogenic and antiapoptotic role of constitutive NF-κB/Rel activity in pancreatic cancer. Int J Cancer 105:735–746. doi:10.1002/ijc.11081
McDonald ER, El-Deiry WS (2000) Cell cycle control as a basis for cancer drug development. Int J Oncol 16:871–886
Mu D, Zhang W, Chu D, Liu T, Xie Y, Fu E, Jin F (2008) The role of calcium, P38 MAPK in dihydroartemisinin-induced apoptosis of lung cancer PC-14 cells. Cancer Chemother Pharmacol 61:639–645. doi:10.1007/s00280-007-0517-5
Nam W, Tak J, Ryu JK, Jung M, Yook JI, Kim HJ, Cha IH (2007) Effects of artemisinin and its derivatives on growth inhibition and apoptosis of oral cancer cells. Head Neck 29:335–340. doi:10.1002/hed.20524
Owa T, Yoshino H, Yoshimatsu K, Nagasu T (2001) Cell cycle regulation in the G1 phase: a promising target for the development of new chemotherapeutic anticancer agents. Curr Med Chem 8:1487–1503
Sherr CJ (1996) Cancer cell cycles. Science 274:1672–1677. doi:10.1126/science.274.5293.1672
Sherr CJ (2004) Principles of tumor suppression. Cell 116:235–246. doi:10.1016/S0092-8674(03)01075-4
Sherr CJ, Roberts JM (1999) CDK inhibitors: positive and negative regulators of G1-phase progression. Genes Dev 13:1501–1512
Sherr CJ, Roberts JM (2004) Living with or without cyclins and cyclin-dependent kinases. Genes Dev 18:2699–2711. doi:10.1101/gad.1256504
Siedle B, Garcia-Pineres AJ, Murillo R, Schulte-Monting J, Castro V, Rungeler P, Klaas CA, Da Costa FB, Kisiel W, Merfort I (2004) Quantitative structure–activity relationship of sesquiterpene lactones as inhibitors of the transcription factor NF-kappaB. J Med Chem 47:6042–6054. doi:10.1021/jm049937r
Singh NP, Lai H (2001) Selective toxicity of dihydroartemisinin and holotransferrin toward human breast cancer cells. Life Sci 70:49–56. doi:10.1016/S0024-3205(01)01372-8
Singh NP, Lai HC (2004) Artemisinin induces apoptosis in human cancer cells. Anticancer Res 24:2277–2280
Singh NP, Lai HC (2005) Synergistic cytotoxicity of artemisinin and sodium butyrate on human cancer cells. Anticancer Res 25:4325–4331
Swanton C (2004) Cell-cycle targeted therapies. Lancet Oncol 5:27–36. doi:10.1016/S1470-2045(03)01321-4
Van Antwerp DJ, Martin SJ, Kafri T, Green DR, Verma IM (1996) Suppression of TNF-α-induced apoptosis by NF-κB. Science 274:787–789. doi:10.1126/science.274.5288.787
van den Heuvel S, Harlow E (1993) Distinct roles for cyclin-dependent kinases in cell cycle control. Science 262:2050–2054. doi:10.1126/science.8266103
Vermeulen K, Berneman ZN, Van Bockstaele DR (2003) The cell cycle: a review of regulation, deregulation and therapeutic targets in cancer. Cell Prolif 36:165–175
Wang W, Abbruzzese JL, Evans DB, Larry L, Cleary KR, Chiao PJ (1999) The nuclear factor-κB RelA transcription factor is constitutively activated in human pancreatic adenocarcinoma cells. Clin Cancre Res 5:119–127
Wang J, Guo Y, Zhang BC, Chen ZT, Gao JF (2007) Induction of apoptosis and inhibition of cell migration and tube-like formation by dihydroartemisinin in murine lymphatic endothelial cells. Pharmacology 80:207–218. doi:10.1159/000104418
Wu XH, Zhou HJ, Lee J (2006) Dihydroartemisinin inhibits angiogenesis induced by multiple myeloma RPMI8226 cells under hypoxic conditions via downregulation of vascular endothelial growth factor expression and suppression of vascular endothelial growth factor secretion. Anticancer Drugs 17:839–848. doi:10.1097/01.cad.0000224443.85834.32
Xiong HQ, Abbruzzese JL, Lin E, Wang L, Zheng L, Xie K (2004) NF-κB activity blockade impairs the angiogenic potential of human pancreatic cancer cells. Int J Cancer 108:181–188. doi:10.1002/ijc.11562
Yamamoto Y, Gaynor RB (2001) Therapeutic potential of inhibition of the NF-κB pathway in the treatment of inflammation and cancer. J Clin Invest 107:135–142. doi:10.1172/JCI11914
Yebra M, Filardo EJ, Bayna EM, Kawahara E, Becker JC, Cheresh DA (1995) Induction of carcinoma cell migration on vitronectin by NF-κB-dependent gene expression. Mol Biol Cell 6:841–850
Acknowledgments
This work was supported in part by Grants from Program for New Century Excellent Talents in University of Chinese Ministry of Education (NCET-07-0248), Science Foundation for Excellent Youth of Heilongjiang Province, China (JC200717), the National Natural Scientific Foundation of China (30901437, 30972907).
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Chen, H., Sun, B., Wang, S. et al. Growth inhibitory effects of dihydroartemisinin on pancreatic cancer cells: involvement of cell cycle arrest and inactivation of nuclear factor-κB. J Cancer Res Clin Oncol 136, 897–903 (2010). https://doi.org/10.1007/s00432-009-0731-0
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DOI: https://doi.org/10.1007/s00432-009-0731-0