Skip to main content

Advertisement

SpringerLink
Log in

A novel antitubulin agent, DPQZ, induces cell apoptosis in human oral cancer cells through Ras/Raf inhibition and MAP kinases activation

  • Molecular Toxicology
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

6-(N,N-Dimethylamino)-2-(naphthalene-1-yl)-4-quinazolinone (DPQZ)-induced apoptosis was accompanied by the characteristics of DNA fragmentation and phosphatidylserine externalization in human oral cancer HSC-3 cells. The IC50 (half maximal inhibitory concentration) value of DPQZ is about 0.25 μM at 24 h. The interference in the dynamics of tubulin and cell division of DPQZ, like vinblastine (0.01 μM), has been proven in this study. Treatment of HSC-3 cells with DPQZ resulted in many of mitotic cells with multipolar spindles. Up-regulation of MAP kinases, such as ERK, JNK, and p38, mediated by DPQZ appears to be involved in DPQZ-induced apoptosis in HSC-3 cells. It is worthy of note that the expression of Ras and c-Raf that lie upstream of ERK were inhibited by DPQZ. In addition, the DPQZ-induced cell death was attenuated by JNK inhibitor SP600125 (3 or 10 μM), not by the ERK or p38 inhibitors. JNK inhibitor abolished the DPQZ-induced increase in the phosphorylation of Bcl-2 and the protein levels of proform caspase-3, caspase-8, and caspase-9, indicating that JNK is an upstream activator of Bcl-2 and caspase family members and plays a key role in DPQZ-induced HSC-3 cell apoptosis. We also attempted to develop an anticancer drug that is designed to kill rapidly dividing cancer cells while causing less damage to normal cells. The DPQZ-induced cytotoxicity against human gingival fibroblasts was less than that against HSC-3 cells. Our work provides a new strategy and mechanism for developing anticancer drug and may contribute to clinical anticancer drug discovery and application.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

DAPI:

4′,6-Diamidino-2-phenylindole dihydrochloride

DMSO:

Dimethylsulfoxide

DPQZ:

6-(N,N-Dimethylamino)-2-(naphthalene-1-yl)-4-quinazolinone

ERK:

Extracellular signal-regulated kinase

FITC:

Fluorescein isothiocyanate

HRP:

Horseradish peroxidase

HSC-3:

Human oral cancer cell line

JNK:

c-Jun N-terminal kinase

MAP kinase:

Mitogen-activated protein kinase

PI:

Propidium iodide

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

References

  • Agarwal R, Kaye SB (2003) Ovarian cancer: strategies for overcoming resistance to chemotherapy. Nat Rev Cancer 3:502–516

    Article  PubMed  CAS  Google Scholar 

  • Amarasinghe HK, Usgodaarachchi US, Johnson NW, Lalloo R, Warnakulasuriya S (2010) Betel-quid chewing with or without tobacco is a major risk factor for oral potentially malignant disorders in Sri Lanka: a case-control study. Oral Oncol 46:297–301

    Article  PubMed  Google Scholar 

  • Basu A, You SA, Haldar S (2000) Regulation of Bcl2 phosphorylation by stress response kinase pathway. Int J Oncol 16:497–500

    PubMed  CAS  Google Scholar 

  • Burns CJ, Fantino E, Phillips ID, Su S, Harte MF, Bukczynska PE, Frazzetto M, Joffe M, Kruszelnicki I, Wang B, Wang Y, Wilson N, Dilley RJ, Wan SS, Charman SA, Shackleford DM, Fida R, Malcontenti-Wilson C, Wilks AF (2009) CYT997: a novel orally active tubulin polymerization inhibitor with potent cytotoxic and vascular disrupting activity in vitro and in vivo. Mol Cancer Ther 8:3036–3045

    Article  PubMed  CAS  Google Scholar 

  • Cao X, Zhang Y, Zou L, Xiao H, Chu Y, Chu X (2010) Persistent oxygen-glucose deprivation induces astrocytic death through two different pathways and calpain-mediated proteolysis of cytoskeletal proteins during astrocytic oncosis. Neurosci Lett 479:118–122

    Article  PubMed  CAS  Google Scholar 

  • Castedo M, Perfettini JL, Roumier T, Andreau K, Medema R, Kroemer G (2004) Cell death by mitotic catastrophe: a molecular definition. Oncogene 23:2825–2837

    Article  PubMed  CAS  Google Scholar 

  • Doyle LA, Ross DD (2003) Multidrug resistance mediated by the breast cancer resistance protein BCRP (ABCG2). Oncogene 22:7340–7358

    Article  PubMed  Google Scholar 

  • Farghaly AM, Mohsen A, Omar ME, Khalil MA, Gaber MA (1990) Synthesis of 3-aryl-2-substituted-4(3H)-quinazolines as potential antimicrobial agents. Farmaco 45:431–438

    PubMed  CAS  Google Scholar 

  • Furukawa Y, Iwase S, Kikuchi J, Terui Y, Nakamura M, Yamada H, Kano Y, Matsuda M (2000) Phosphorylation of Bcl-2 protein by CDC2 kinase during G2/M phases and its role in cell cycle regulation. J Biol Chem 275:21661–21667

    Article  PubMed  CAS  Google Scholar 

  • Gottesman MM, Fojo T, Bates SE (2002) Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer 2:48–58

    Article  PubMed  CAS  Google Scholar 

  • Gupta B, Ariyawardana A, Johnson NW (2012) The epidemic of oral cancer in India continues unabated: need for new policy initiatives. Oral Oncol 48:e31–e32

    Article  PubMed  Google Scholar 

  • Hadfield JA, Ducki S, Hirst N, McGown AT (2003) Tubulin and microtubules as targets for anticancer drugs. Prog Cell Cycle Res 5:309–325

    PubMed  Google Scholar 

  • Haldar S, Basu A, Croce CM (1997) Bcl2 is the guardian of microtubule integrity. Cancer Res 57:229–233

    PubMed  CAS  Google Scholar 

  • Kartalou M, Essigmann JM (2001) Mechanisms of resistance to cisplatin. Mutat Res 478:23–43

    Article  PubMed  CAS  Google Scholar 

  • Kyriakis JM, App H, Zhang XF, Banerjee P, Brautigan DL, Rapp UR, Avruch J (1992) Raf-1 activates MAP kinase–kinase. Nature 358:417–421

    Article  PubMed  CAS  Google Scholar 

  • Lai MY, Hour MJ, Leung WC, Yang WH, Lee HZ (2007) Chaperones are the target in aloe-emodin-induced human lung nonsmall carcinoma H460 cell apoptosis. Eur J Pharmacol 573:1–10

    Article  PubMed  CAS  Google Scholar 

  • Lee HZ, Yang WH, Hour MJ, Wu CY, Peng WH, Bao BY, Han PH, Bau DT (2010) Photodynamic activity of aloe-emodin induces resensitization of lung cancer cells to anoikis. Eur J Pharmacol 648:50–58

    Article  PubMed  CAS  Google Scholar 

  • Lin CC, Yang JS, Chen JT, Fan S, Yu FS, Yang JL, Lu CC, Kao MC, Huang AC, Lu HF, Chung JG (2007) Berberine induces apoptosis in human HSC-3 oral cancer cells via simultaneous activation of the death receptor-mediated and mitochondrial pathway. Anticancer Res 27:3371–3378

    PubMed  CAS  Google Scholar 

  • Ling YH, Tornos C, Perez-Soler R (1998) Phosphorylation of Bcl-2 is a marker of M phase events and not a determinant of apoptosis. J Biol Chem 273:18984–18991

    Article  PubMed  CAS  Google Scholar 

  • Liu Y, Guyton KZ, Gorospe M, Xu Q, Lee JC, Holbrook NJ (1996) Differential activation of ERK, JNK/SAPK and p38/CSBP/RK map kinase family members during the cellular response to arsenite. Free Radic Biol Med 21:771–781

    Article  PubMed  CAS  Google Scholar 

  • Ma D, Warabi E, Yanagawa T, Kimura S, Harada H, Yamagata K, Ishii T (2009) Peroxiredoxin I plays a protective role against cisplatin cytotoxicity through mitogen activated kinase signals. Oral Oncol 45:1037–1043

    Article  PubMed  CAS  Google Scholar 

  • Manfredi JJ, Parness J, Horwitz SB (1982) Taxol binds to cellular microtubules. J Cell Biol 94:688–696

    Article  PubMed  CAS  Google Scholar 

  • Modh RP, Patel AC, Chikhalia KH (2012) Design, synthesis & biological evaluation of some novel quinazolinone scaffolds. Med Chem 8:182–192

    PubMed  CAS  Google Scholar 

  • Mohamed MS, Kamel MM, Kassem EM, Abotaleb N, Khedr M, Ahmed MF (2011) Synthesis, biological evaluation and molecular docking of quinazoline-4(1H)-one derivatives as anti-inflammatory and analgesic agents. Acta Pol Pharm 68:665–675

    PubMed  CAS  Google Scholar 

  • Mollinedo F, Gajate C (2003) Microtubules, microtubule-interfering agents and apoptosis. Apoptosis 8:413–450

    Article  PubMed  CAS  Google Scholar 

  • Pathan N, Aime-Sempe C, Kitada S, Haldar S, Reed JC (2001) Microtubuletargeting drugs induce Bcl-2 phosphorylation and association with Pin1. Neoplasia 3:70–79

    Article  PubMed  CAS  Google Scholar 

  • Peters JM (2002) The anaphase-promoting complex: proteolysis in mitosis and beyond. Mol Cell 9:931–943

    Article  PubMed  CAS  Google Scholar 

  • Raingeaud J, Gupta S, Rogers JS, Dickens M, Han J, Ulevitch RJ, Davis RJ (1995) Pro-inflammatory cytokines and environmental stress cause p38 mitogen-activated protein kinase activation by dual phosphorylation on tyrosine and threonine. J Biol Chem 270:7420–7426

    Article  PubMed  CAS  Google Scholar 

  • Rodriguez-Viciana P, Oses-Prieto J, Burlingame A, Fried M, McCormick F (2006) A phosphatase holoenzyme comprised of Shoc2/Sur8 and the catalytic subunit of PP1 functions as an M-Ras effector to modulate Raf activity. Mol Cell 22:217–230

    Article  PubMed  CAS  Google Scholar 

  • Roninson IB, Broude EV, Chang BD (2001) If not apoptosis, then what? Treatment-induced senescence and mitotic catastrophe in tumor cells. Drug Resist Updat 4:303–313

    Article  PubMed  CAS  Google Scholar 

  • Sánchez-Perez I, Murguía JR, Perona R (1998) Cisplatin induces a persistent activation of JNK that is related to cell death. Oncogene 16:533–540

    Article  PubMed  Google Scholar 

  • Sharma PC, Kaur G, Pahwa R, Sharma A, Rajak H (2011) Quinazolinone analogs as potential therapeutic agents. Curr Med Chem 18:4786–4812

    Article  PubMed  CAS  Google Scholar 

  • Srivastava RK, Mi QS, Hardwick JM, Longo DL (1999) Deletion of the loop region of Bcl-2 completely blocks paclitaxel-induced apoptosis. Proc Natl Acad Sci USA 96:3775–3780

    Article  PubMed  CAS  Google Scholar 

  • Terai K, Matsuda M (2005) Ras binding opens c-Raf to expose the docking site for mitogen-activated protein kinase kinase. EMBO Rep 6:251–255

    Article  PubMed  CAS  Google Scholar 

  • Wu YC, Hour MJ, Leung WC, Wu CY, Liu WZ, Chang YH, Lee HZ (2011) 2-(Naphthalene-1-yl)-6-pyrrolidinyl-4-quinazolinone inhibits skin cancer M21 cell proliferation through aberrant expression of microtubules and the cell cycle. Pharmacol Exp Ther 338:942–951

    Article  CAS  Google Scholar 

  • Yang JS, Hour MJ, Kuo SC, Huang LJ, Lee MR (2004) Selective induction of G2/M arrest and apoptosis in HL-60 by a potent anticancer agent, HMJ-38. Anticancer Res 24:1769–1778

    PubMed  CAS  Google Scholar 

  • Yeh FT, Wu CH, Lee HZ (2003) Signaling pathway for aloe-emodin-induced apoptosis in human H460 lung nonsmall carcinoma cell. Int J Cancer 106:26–33

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Miss Ciou-Wei Jhou for technical assistance and Dr. Yi-Yu Wu for her critical reading of the manuscript. This work was supported by China Medical University Grant CMU99-COL-09 and CMU100-TS-03 of the Republic of China.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. School of Pharmacy, China Medical University, 91, Hsueh-Shih Road, Taichung, 40402, Taiwan

    Mann-Jen Hour & Hong-Zin Lee

  2. Department of Oral Hygiene, College of Dental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Kun-Tsung Lee

  3. School of Chinese Medicine, China Medical University, Taichung, Taiwan

    Yang-Chang Wu

  4. Department of Internal Medicine, Chang-Hua Hospital, Chang-Hua, Taiwan

    Chi-Yu Wu

  5. School of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, China Medical University, Taichung, Taiwan

    Bang-Jau You

  6. Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei, Taiwan

    Tai-Lin Chen

Authors
  1. Mann-Jen Hour
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kun-Tsung Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yang-Chang Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chi-Yu Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Bang-Jau You
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tai-Lin Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hong-Zin Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hong-Zin Lee.

Additional information

Mann-Jen Hour and Kun-Tsung Lee equally contributed to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hour, MJ., Lee, KT., Wu, YC. et al. A novel antitubulin agent, DPQZ, induces cell apoptosis in human oral cancer cells through Ras/Raf inhibition and MAP kinases activation. Arch Toxicol 87, 835–846 (2013). https://doi.org/10.1007/s00204-012-0991-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-012-0991-3

Keywords

Search

Navigation