DOX-MTX-NPs Augment p53 mRNA Expression in OSCC Model in Rat: Effects of IV and Oral Routes

  • Abbasi, Mehran Mesgari (Drug Applied Research Center) ;
  • Khiavi, Monir Moradzadeh (Department of Oral Pathology, Faculty of Dentistry, Tehran University of Medical Sciences, International Campus) ;
  • Monfaredan, Amir (Department of Hematology, Faculty of Medicine, Tabriz Branch, Islamic Azad University) ;
  • Hamishehkar, Hamed (Drug Applied Research Center) ;
  • Seidi, Khaled (Department of Medical Biotechnology, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences) ;
  • Jahanban-Esfahlan, Rana (Student Research Committee)
  • Published : 2014.10.23


Background: Oral squamous cell carcinoma (OSCC) is the sixth most common malignancy worldwide. Cancer development and progression require inactivation of tumor suppressor genes and activation of proto-oncogenes. The well recognized mechanism of action demonstrated for chemotherapeutic agents is induction of apoptosis via reactivation of p53. In this context, we evaluate the efficacy of IV and oral routes of our novel PH and temperature sensitive doxorubicin-methotrexate-loaded nanoparticles (DOX-MTX NP) in affecting p53 profile in an OSCC rat model. Methods: In this study, 120 male rats were divided into 8 groups of 15 animals each. The new formulated DOX-MTX NP and free doxorubicin were IV and orally given to rats with 4-nitroquinoline-1-oxide induced OSCC. Results: Results showed that both DOX and DOX-MTX-NP caused significant increase in mRNA levels of P53 compared to the untreated group (p<0.000). With both DOX and DOX-MTX NP, the IV mode was more effective than the oral (gavage) route (p<0.000). Surprisingly, in oral mode, p53 mRNA was not affected in DOX treated groups (p>0.05), Nonetheless, both IV and oral administration of MTX-DOX NP showed superior activity (~3 fold) over free DOX in reactivation of p53 in OSCC (p<0.000). The effectiveness of oral route in group treated with nanodrug accounts for the enhanced bioavailability of nanoparticulated DOX-MTX compared to free DOX. Moreover, in treated groups, tumor stage was markedly related to the amount of p53 mRNA (p<0.05). Conclusion: Both oral and IV application of our novel nanodrug possesses superior activity over free DOX-in up-regulation of p53 in a OSCC model and this increase in p53 level associated with less aggressive tumors in our study. Although, impressive results obtained with IV form of nanodrug (-21 fold increase in p53 mRNA level) but both forms of nanodrug are effective in OSCC, with less toxicity normal cells.


p53;DOX-MTX-NPs;oral squamous cell carcinoma;oral and IV route;rat model


  1. Benival D, PV D (2012). Lipomer of doxorubicin hydrochloride for enhanced oral bioavailability. Int J Pharm, 423, 554-61.
  2. Abusail M, Dirweesh AM, Salih RA, et al (2013). Expression of EGFR and p53 in Head and Neck Tumors among Sudanese Patients. Asian Pac J Cancer Prev, 14, 6415-8
  3. Bae Y (2010). Drug delivery systems using polymer nanoassemblies for cancer treatment. Ther Deliv, 1, 361-3.
  4. Baykara M, Buyukberber S, Ozturk B, et al (2013). Efficacy and safety of concomitant chemoradiotherapy with cisplatin and docetaxel in patients with locally advanced squamous cell head and neck cancers. Asian Pac J Cancer Prev, 14, 2557-61.
  5. Chen Y, Wan Y, Wang Y, et al (2011). Anticancer efficacy enhancement and attenuation of side effects of doxorubicin with titanium dioxide nanoparticles. Int J Nanomedicine, 6, 2321-6.
  6. Cipriani P, Ruscitti P, Carubbi F, et al (2014). Methotrexate in Rheumatoid Arthritis: Optimizing Therapy Among Different Formulations. Current and Emerging Paradigms. Clin Ther, 36, 427-35.
  7. Deng Y, Zhang H (2013). The synergistic effect and mechanism of doxorubicin-ZnO nanocomplexes as a multimodal agent integrating diverse anticancer therapeutics. Int J Nanomedicine, 8, 1835-41.
  8. Duong HH, Yung LY (2013). Synergistic co-delivery of doxorubicin and paclitaxel using multi-functional micelles for cancer treatment. Int J Pharm, 454, 486-95.
  9. Jahanban Esfahlan R, Zarghami N, Jahanban Esfahlan A, et al (2011a). The possible impact of obesity on androgen, progesterone and estrogen receptors (ERa and ERb) gene expression in breast cancer patients. Breast Cancer, 5, 227-37.
  10. Guhagarkar SA, Gaikwad RV, Samad A, et al (2010). Polyethylene sebacate-doxorubicin nanoparticles for hepatic targeting. Int J Pharm, 401, 113-22.
  11. Hu CM, Aryal S, Zhang L (2010). Nanoparticle-assisted combination therapies for effective cancer treatment. Ther Deliv, 1, 323-34.
  12. Huang WY, Yang PM, Chang YF, et al (2011). Methotrexate induces apoptosis through p53/p21-dependent pathway and increases E-cadherin expression through downregulation of HDAC/EZH2. Biochem Pharmacol, 81, 510-7.
  13. Jahanban Esfahlan R, Zarghami N, Rahmati-Yamchi M, et al (2011b). Quantification of Steroid Receptors Gene Expression in Breast Cancer Patients: Possible Correlation with Serum Level of Adipocytokines. Journal of Cancer Therapy, 2, 659-65.
  14. Jahanban Esfahlan R, Zarghami N, Valiyari S, et al (2012). Adiponectin Can Affect ER Signaling in Obese Breast Cancer Patients. Journal of Cancer Therapy, 3, 115-21
  15. Jain S, Patil SR, Swarnakar NK, et al (2012). Oral delivery of doxorubicin using novel polyelectrolyte-stabilized liposomes (layersomes). Mol Pharm, 9, 2626-35.
  16. Jones KR, Lodge-Rigal RD, Reddick RL, et al (1992). Prognostic factors in the recurrence of stage I and II squamous cell cancer of the oral cavity. Arch Otolaryngol Head Neck Surg, 118, 483-5.
  17. Kademani D, Bell RB, Bagheri S, et al (2005). Prognostic factors in intraoral squamous cell carcinoma: the influence of histologic grade. J Oral Maxillofac Surg, 63, 1599-605.
  18. Liboiron B, Mayer L (2014). Nanoscale particulate systems for multidrug delivery: towards improved combination chemotherapy. Ther Deliv, 5, 149-71.
  19. Kalaria DR, Sharma G, Beniwal V, et al (2009). Design of biodegradable nanoparticles for oral delivery of doxorubicin: in vivo pharmacokinetics and toxicity studies in rats. Pharm Res, 26, 492-501.
  20. Kim HR, Christensen R, Park NH, et al (2001). Elevated expression of hTERT is associated with dysplastic cell transformation during human oral carcinogenesis in situ. Clin Cancer Res, 7, 3079-86.
  21. Lasrado S, Moras K, GJ P, et al (2014). Role of concomitant chemoradiation in locally advanced head and neck cancers. Asian Pac J Cancer Prev, 15, 4147-52.
  22. Lippman SM, Hong WK (2001). Molecular markers of the risk of oral cancer. N Engl J Med, 344, 1323-6.
  23. Liu L, Zhang D, Jiao JH, et al (2014). Association between the TP53BP1 rs2602141 A/C polymorphism and cancer risk: a systematic review and meta-analysis. Asian Pac J Cancer Prev, 15, 2917-22.
  24. Massano J, Regateiro FS, Januario G, et al (2006). Oral squamous cell carcinoma: review of prognostic and predictive factors. Oral Surg.Oral Med.Oral Pathol.Oral Radiol Endod, 102, 67-76.
  25. Mehdipour M, Taghavi ZA, Mesgari AM, et al (2013). Evaluation of the Effect of Two Systemic Doses of HESA-A on Prevention of Induced Tongue Neoplasm in Rats. J Dent Res Dent Clin Dent Prospects, 7, 218-24.
  26. Mesgari Abbasi M, Monfaredan A, Hamishehkar H, et al (2014). Novel DOX-MTX NPs improve the OSCC clinical outcome by down regulation of lymph dissemination factor VEGF-C expression in vivo: effect of oral and IV modalities. Asian Pac J Cancer Prev, 15 (15), 6227-32.
  27. Salehi R, Hamishehkar H, Eskandani M, et al (2014). Development of dual responsive nanocomposite for simultaneous delivery of anticancer drugs. J.Drug Target.
  28. Mollazade M, Nejati-Koshki K, Akbarzadeh A, et al (2013). PAMAM dendrimers augment inhibitory effects of curcumin on cancer cell proliferation: possible inhibition of telomerase. Asian Pac J Cancer Prev, 14, 6925-8.
  29. Montoro JR, Ricz HA, Souza L, et al (2008). Prognostic factors in squamous cell carcinoma of the oral cavity. Braz J Otorhinolaryngol, 74, 861-6.
  30. Nasiri M, Zarghami N, Nejati Koshki K, et al (2013). Curcumin and Silibinin Inhibit Telomerase Expression in T47D Human Breast Cancer Cells. Asian Pac J Cancer Prev, 14, 3449-53
  31. Rossi B, Schinzari G, Maccauro G, et al (2010). Neoadjuvant multidrug chemotherapy including high-dose methotrexate modifies VEGF expression in osteosarcoma: an immunohistochemical analysis. BMC Musculoskelet Disord, 11, 34.
  32. Schliephake H (2003). Prognostic relevance of molecular markers of oral cancer-a review. Int J Oral Maxillofac Surg, 32, 233-45.
  33. Tacar O, Sriamornsak P, Dass CR (2013). Doxorubicin: an update on anticancer molecular action, toxicity and novel drug delivery systems. J Pharm Pharmacol, 65, 157-70.
  34. Valiyari S, Jahanban-Esfahlan R, Zare Shahneh F, et al (2013). Cytotoxic and apoptotic activity of Scrophularia oxysepala in MCF-7 human breast cancer cells. Toxicological & Environmental Chemistry, 95, 1208-20.
  35. Wang Y, Wei X, Zhang C, et al (2010). Nanoparticle delivery strategies to target doxorubicin to tumor cells and reduce side effects. Ther Deliv, 1, 273-87.

Cited by

  1. Oral and IV Dosages of Doxorubicin-Methotrexate loaded-Nanoparticles Inhibit Progression of Oral Cancer by Down-Regulation of Matrix Methaloproteinase 2 Expression in Vivo vol.15, pp.24, 2015,
  2. Hesa-A Improves Clinical Outcome of Oral Carcinoma by Affecting p53 Gene Expression in vivo vol.16, pp.10, 2015,
  3. Anti Tumoral Properties of Punica Granatum (Pomegranate) Peel Extract on Different Human Cancer Cells vol.16, pp.14, 2015,
  4. Anti Proliferative Properties of Melissa officinalis in Different Human Cancer Cells vol.16, pp.14, 2015,
  5. Hesa-A Down-Regulates erb/b2 Oncogene Expression and Improves Outcome of Oral Carcinoma in a Rat Model vol.16, pp.16, 2015,
  6. Anti-Proliferative Effects of Hesa-A on Human Cancer Cells with Different Metastatic Potential vol.16, pp.16, 2015,
  7. Anti Tumoral Properties of Punica granatum (Pomegranate) Seed Extract in Different Human Cancer Cells vol.17, pp.3, 2016,
  8. Modulating tumor hypoxia by nanomedicine for effective cancer therapy vol.233, pp.3, 2017,
  9. Combination of nanotechnology with vascular targeting agents for effective cancer therapy vol.233, pp.4, 2017,