New formulated "DOX-MTX-loaded Nanoparticles" Down-regulate HER2 Gene Expression and Improve the Clinical Outcome in OSCCs Model in Rat: the Effect of IV and Oral Modalities

  • Published : 2014.11.28


Background: Oral squamous cell carcinoma (OSCC) remains as one of the most difficult malignancies to control because of its high propensity for local invasion and cervical lymph node dissemination. In this study, we evaluate the efficacy of our novel pH and temperature sensitive doxorubicin-methotrexate-loaded nanoparticles (DOX-MTX NP) in affecting HER2 expression profile in OSCC model in rat. Results: DOX-MTX- nanoparticle complexes caused significant decrease in mRNA level of HER2 compared to untreated cancers (p<0.05) and this finding was more pronounced with the IV mode (p<0.000). Surprisingly, HER2 mRNA was not affected in DOX treated as compared to the control group (p>0.05). On the other hand, in the DOX-MTX NP treated group, fewer tumors characterized with advanced stage and decreased HER2 paralleled improved clinical outcome (P<0.05). Moreover, the effectiveness of the oral route in the group treated with nanodrug accounted for the enhanced bioavailability of nanoparticulated DOX-MTX compared to free DOX. Furthermore, there was no significant difference in mRNA level of HER2 (p>0.05). Conclusions: Influence of HER2 gene expression is a new feature and mechanism of action observed only in dual action DOX-MTX-NPs treated groups. Down-regulation of HER2 mRNA as a promising marker and prognosticator of OSCC adds to the cytotoxic benefits of DOX in its new formulation. Both oral and IV application of this nanodrug could be used, with no preferences in term of their safety or toxicity. As HER2 is expressed abundantly by a wide spectrum of tumors, i DOX-MTX NPs may be useful for a wide-spectrum of lesions. However, molecular mechanisms underlying HER2 down regulation induced by DOX-MTX NPs remain to be addressed.


  1. 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.
  2. 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.
  3. Salehi R, Hamishehkar H, Eskandani M, et al (2014). Development of dual responsive nanocomposite for simultaneous delivery of anticancer drugs. J Drug Target, 22, 327-42.
  4. Sardari Y, Pardis S, Tadbir AA, et al (2012). HER2/neu expression in head and neck squamous cell carcinoma patients is not significantly elevated. Asian Pac J Cancer Prev, 13, 2891-6.
  5. Schliephake H (2003). Prognostic relevance of molecular markers of oral cancer--a review. Int J Oral Maxillofac Surg, 32, 233-45.
  6. 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.
  7. 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.
  8. Yoo HS, Park TG (2004). Folate-receptor-targeted delivery of doxorubicin nano-aggregates stabilized by doxorubicin-PEG-folate conjugate. J Control Release, 100, 247-56.
  9. 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.
  10. Bae Y (2010). Drug delivery systems using polymer nanoassemblies for cancer treatment. Ther Deliv, 1, 361-3.
  11. 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.
  12. Bell R, Kademani D, Homer L, et al (2007). Tongue cancer: Is there a difference in survival compared with other subsites in the oral cavity? J Oral Maxillofac Surg, 65, 229-36.
  13. Benival D, PV D (2012). Lipomer of doxorubicin hydrochloride for enhanced oral bioavailability. Int J Pharm, 423, 554-61.
  14. Brandwein-Gensler M, Teixeira MS, Lewis CM, et al (2005). Oral squamous cell carcinoma: histologic risk assessment, but not margin status, is strongly predictive of local diseasefree and overall survival. Am J Surg Pathol, 29, 167-78.
  15. 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.
  16. 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.
  17. 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.
  18. Gibson RJ, Bowen JM, Cummins AG, et al (2005). Relationship between dose of methotrexate, apoptosis, p53/p21 expression and intestinal crypt proliferation in the rat. Clin Exp Med, 4, 188-95.
  19. Hu CM, Aryal S, Zhang L (2010). Nanoparticle-assisted combination therapies for effective cancer treatment. Ther Deliv, 1, 323-34.
  20. 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.
  21. 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 (Auckl), 227-37.
  22. 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. J Cancer Therapy, 2, 659-65.
  23. Jahanban Esfahlan R, Zarghami N, Valiyari S, et al (2012). Adiponectin can affect er signaling in obese breast cancer patients. J Cancer Therapy, 3, 115-21.
  24. 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.
  25. 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.
  26. 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.
  27. Khademi B, Khademi B, Ghaderi A, et al (2013). Early Detection of Serum Levels of HER-2 in Patients with Head and Neck Squamous Cell Carcinoma. Iran J Otorhinolaryngol, 25, 161-8.
  28. 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.
  29. 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.
  30. Li B, Wang M, Wen Y, et al (1992). [erb-B oncogene in human oral squamous cell carcinomas]. Hua Xi Yi Ke Da Xue Xue Bao, 23, 284-7.
  31. Lippman SM, Hong WK (2001). Molecular markers of the risk of oral cancer. N Engl J Med, 344, 1323-6.
  32. 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.
  33. 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.
  34. Mesgari Abbasi M, Monfaredan A, Hamishehkar H, et al (2014b). DOX-MTX-NPs augment p53 mRNA expression in OSCCs model in rat: the effect of IV and oral dosages. Asian Pac J Cancer Prev, 19, 8377-82.
  35. Mesgari Abbasi M, Monfaredan A, Hamishehkar H, et al (2014d). Novel DOX-MTX nanoparticles improve oral SCC clinical outcome by down regulation of lymph dissemination factor VEGF-C expression in vivo: oral and IV modalities. Asian Pac J Cancer Prev, 15, 6227-32.

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