Cancer Chemotherapy and Pharmacology

, Volume 61, Issue 4, pp 639–645 | Cite as

The role of calcium, P38 MAPK in dihydroartemisinin-induced apoptosis of lung cancer PC-14 cells

  • Deguang Mu
  • Wei Zhang
  • Dongling Chu
  • Tonggang Liu
  • Yonghong Xie
  • Enqing Fu
  • Faguang Jin
Original Article

Abstract

Introduction

Dihydroartemisinin (DHA), a semi-synthetic derivative of artemisinin isolated from the traditional Chinese herb Artemisia annua, is an effective novel antimalarial drug. Recent studies suggest that it also has anticancer effect.

Purpose

The present study was designed to investigate the effects of DHA on cultured human lung cancer cells (PC-14 cells) to better understand its apoptosis and apoptosis-related factors in vitro.

Methods

The cell viability was measured by MTT assay. The apoptosis induction was examined by DNA ladder and flow cytometry. The intracellular-free calcium concentration in the lung cancer cells were evaluated by laser scanning confocal microscopy with Fura-3/AM as probe. The associated gene expression was examined by Western blot.

Results

After treatment with DHA, a decrease in the viability of PC-14 cells and apoptosis were observed. DHA-induced apoptosis were accompanied by an increase of Ca2+ and activation of p38. Deleted levels of Ca2+ by BAPTA-AM 20 μM or inhibition of p38 by its selective inhibitor SB202190 then led to decreased DHA-induced apoptosis.

Conclusions

These results demonstrated that DHA can induce apoptosis of lung cancer cell line PC-14 cells and calcium and p38 play important roles in the apoptotic signalling pathways.

Keywords

Dihydroartemisinin PC-14 cell Intracellular calcium concentration p38 MAPK Apoptosis 

References

  1. 1.
    Jacobson MD, Weil M, Raff MC (1997) Programmed cell death in animal development. Cell 88(3):347–354PubMedCrossRefGoogle Scholar
  2. 2.
    Fadeel B, Orrenius S, Zhivotovsky B (1999) Apoptosis in human disease: a new skin for the old ceremony? Biochem Biophys Res Commun 266(3):699–717PubMedCrossRefGoogle Scholar
  3. 3.
    Gerl R, Vaux DL (2005) Apoptosis in the development and treatment of cancer. Carcinogenesis 26(2):263–270PubMedCrossRefGoogle Scholar
  4. 4.
    Gordi T, Lepist EI (2004) Artemisinin derivatives: toxic for laboratory animals, safe for humans? Toxicol Lett 147(2):99–107PubMedCrossRefGoogle Scholar
  5. 5.
    Singh NP, Lai HC (2004) Artemisinin induces apoptosis in human cancer cells. Anticancer Res 24(4):2277–2280PubMedGoogle Scholar
  6. 6.
    Efferth T, Dunstan H, Sauerbrey A, Miyachi H, Chitambar CR (2001) The anti-malarial artesunate is also active against cancer. Int J Oncol 18(4):767–773PubMedGoogle Scholar
  7. 7.
    Berger TG, Dieckmann D, Efferth T, Schultz ES, Funk JO, Baur A, Schuler G (2005) Artesunate in the treatment of metastatic uveal melanoma–first experiences. Oncol Rep 14(6):1599–1603PubMedGoogle Scholar
  8. 8.
    Susin SA, Zamzami N, Kroemer G (1998) Mitochondria as regulators of apoptosis: doubt no more. Biochim Biophys Acta 1366(1–2):151–165PubMedGoogle Scholar
  9. 9.
    Nicotera P, Orrenius S (1998) The role of calcium in apoptosis. Cell Calcium 23(2–3):173–180PubMedCrossRefGoogle Scholar
  10. 10.
    Zhou YQ, Wang B, Mark E, Chung DH (2006) Oxidative stress-induced intestinal epithelial cell apoptosis is mediated by p38 MAPK. Biochem Biophys Res Commun 350(4):860–865PubMedCrossRefGoogle Scholar
  11. 11.
    Lee SK, Jang HJ, Lee HJ, Lee J, Jeon BH, Jun CD, Lee SK, Kim EC (2006) p38 and ERK MAP kinase mediates iron chelator-induced apoptosis and -suppressed differentiation of immortalized and malignant human oral keratinocytes. Life Sci 79(15):1419–1427PubMedCrossRefGoogle Scholar
  12. 12.
    Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65(1–2):55–63PubMedCrossRefGoogle Scholar
  13. 13.
    Kotamraju S, Konorev EA, Joseph J, Kalyanaraman B (2000) Doxorubicin-induced apoptosis in endothelial cells and cardiomyocytes is ameliorated by nitrone spin traps and ebselen. Role of reactive oxygen and nitrogen species. J Biol Chem 275(43):33585–33592PubMedCrossRefGoogle Scholar
  14. 14.
    Efferth T, Olbrich A, Bauer R (2002) mRNA expression profiles for the response of human tumor cell lines to the antimalarial drugs artesunate, arteether, and artemether. Biochem Pharmacol 64(4):617–623PubMedCrossRefGoogle Scholar
  15. 15.
    Huan-huan C, Li-Li Y, Shang-Bin L (2004) Artesunate reduces chicken chorioallantoic membrane neovascularisation and exhibits antiangiogenic and apoptotic activity on human microvascular dermal endothelial cell. Cancer Lett 211(2):163–173PubMedCrossRefGoogle Scholar
  16. 16.
    Efferth T, Benakis A, Romero MR, Tomicic M, Rauh R, Steinbach D, Hafer R, Stamminger T, Oesch F, Kaina B, Marschall M (2004) Enhancement of cytotoxicity of artemisinins toward cancer cells by ferrous iron. Free Radic Biol Med 37(7):998–1009PubMedCrossRefGoogle Scholar
  17. 17.
    Wu GD, Zhou HJ, Wu XH (2004) Apoptosis of human umbilical vein endothelial cells induced by artesunate. Vascul Pharmacol 41(6):205–212PubMedCrossRefGoogle Scholar
  18. 18.
    Zheng GQ (1994) Cytotoxic terpenoids and flavonoids from Artemisia annua. Planta Med 60(1):54–57PubMedCrossRefGoogle Scholar
  19. 19.
    Beekman AC, Barentsen AR, Woerdenbag HJ, Van Uden W, Pras N, Konings AW, el-Feraly FS, Galal AM, Wikstrom HV (1997) Stereochemistry-dependent cytotoxicity of some artemisinin derivatives. J Nat Prod 60(4):325–330PubMedCrossRefGoogle Scholar
  20. 20.
    Beekman AC, Woerdenbag HJ, Van Uden W, Pras N, Konings AW, Wikstrom HV (1997) Stability of artemisinin in aqueous environments: impact on its cytotoxic action to Ehrlich ascites tumour cells. J Pharm Pharmacol 49(12):1254–1258PubMedGoogle Scholar
  21. 21.
    Venkatachalam K, van Rossum DB, Patterson RL, Ma HT, Gill DL (2002) The cellular and molecular basis of store-operated calcium entry. Nat Cell Biol 4(11):E263–E272PubMedCrossRefGoogle Scholar
  22. 22.
    McConkey DJ, Orrenius S (1997) The role of calcium in the regulation of apoptosis. Biochem Biophys Res Commun 239(2):357–366PubMedCrossRefGoogle Scholar
  23. 23.
    Grethe S, Coltella N, Di Renzo MF, Porn-Ares MI (2006) p38 MAPK downregulates phosphorylation of Bad in doxorubicin-induced endothelial apoptosis. Biochem Biophys Res Commun 347(3):781–790PubMedCrossRefGoogle Scholar
  24. 24.
    Jiang DJ, Jia SJ, Dai Z, Li YJ (2006) Asymmetric dimethylarginine induces apoptosis via p38 MAPK/caspase-3-dependent signaling pathway in endothelial cells. J Mol Cell Cardiol 40(4):529–539PubMedCrossRefGoogle Scholar
  25. 25.
    Petersen C, Svechnikov K, Froysa B, Soder O (2005) The p38 MAPK pathway mediates interleukin-1-induced Sertoli cell proliferation. Cytokine 32(1):51–59PubMedCrossRefGoogle Scholar
  26. 26.
    Senokuchi T, Matsumura T, Sakai M, Matsuo T, Yano M, Kiritoshi S, Sonoda K, Kukidome D, Nishikawa T, Araki E (2004) Extracellular signal-regulated kinase and p38 mitogen-activated protein kinase mediate macrophage proliferation induced by oxidized low-density lipoprotein. Atherosclerosis 176(2):233–245PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Deguang Mu
    • 1
  • Wei Zhang
    • 2
  • Dongling Chu
    • 1
  • Tonggang Liu
    • 1
  • Yonghong Xie
    • 1
  • Enqing Fu
    • 1
  • Faguang Jin
    • 1
  1. 1.Department of Respiratory Disease, Tangdu HospitalFourth Military Medical UniversityXi’an cityChina
  2. 2.Department of MicrobiologyFourth Military Medical UniversityXi’an cityChina

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