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Inhibition of tumor growth by recombinant adenovirus containing human lactoferrin through inducing tumor cell apoptosis in mice bearing EMT6 breast cancer

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Abstract

Human lactoferrin (hLTF), an 80-kDa iron-binding glycoprotein, has antitumor activity. In this study, a recombinant adenovirus containing the human lactoferrin cDNA (ad-rhLTF) was constructed and its effect on tumor growth was investigated in mice bearing EMT6 breast cancer. Ad-rhLTF was injected seven times within 14 days into the tumor site at two concentrations (108 and 5 × 108 pfu/mL) in mice bearing EMT6 breast cancer. Injected ad-rhLTF had considerable cytotoxicity on mice breast cancer, and significantly reducing the weight of tumor produced and increasing the tumor inhibition rate up to 52.64%. The presence of apoptotic cells was confirmed using TUNEL staining and flow cytometry assays. At the same time, RTPCR and Western blot analyses demonstrated that ad-rhLTF also decreased expression of Bcl-2 and increased Bax and caspase 3 expressions. Therefore, we conclude that ad-rhLTF inhibits tumor growth by inducing tumor cell apoptosis in mice with breast cancer by triggering the mitochondrial-dependent pathway and activation of caspase 3. The results indicate that ad-rhLTF might be a promising drug for breast cancer gene therapy.

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References

  • Artym, J., Zimecki, M., Paprocka, M., and Kruzel, M. L., Orally administered lactoferrin restores humoral immune response in immunocompromised mice. Immunol. Lett., 89, 9–15 (2003).

    Article  PubMed  CAS  Google Scholar 

  • Cerven, D., DeGeorge, G., and Bethell, D., 28-Day repeated dose oral toxicity of recombinant human holo-lactoferrin in rats. Regul. Toxicol. Pharmacol., 52, 174–179 (2008).

    Article  PubMed  CAS  Google Scholar 

  • Cheung, T. H., Chung, T. K., Lo, K. W., Yu, M. Y., Krajewski, S., Reed, J. C., and Wong, Y. F., Apoptosis-related proteins in cervical intraepithelial neoplasia and squamous cell carcinoma of the cervix. Gynecol. Oncol., 86, 14–18 (2002).

    Article  PubMed  Google Scholar 

  • Conneely, O. M., Anti-inflammatory activities of lactoferrin. J. Am. Coll. Nutr., 20, 389s–395s (2001).

    PubMed  CAS  Google Scholar 

  • Fujita, K., Matsuda, E., Sekine, K., Ligo, M., and Tsuda, H., Lactoferrin enhances Fas expression and apoptosis in the colon mucosa of azoxymethane-treated rats. Carcinogenesis, 25, 1961–1966 (2004).

    Article  PubMed  CAS  Google Scholar 

  • González-Chávez, S. A., Arévalo-Gallegos, S., and Rascón-Cruz, Q., Lactoferrin: structure, function and applications. Int. J. Antimicrob. Agents, 33, 301.e1–301.e8 (2009).

    Article  Google Scholar 

  • Han, Z. S., Li, Q. W., Zhang, Z. Y., Xiao, B., Gao, D. W., Wu, S. Y., Li, J., Zhao, H. W., Jiang, Z. L., and Hu, J. H., Highlevel expression of human lactoferrin in the milk of goats by using replication-defective adenoviral vectors. Protein Expr. Purif., 53, 225–231 (2007).

    Article  PubMed  CAS  Google Scholar 

  • Han, Z. S., Li, Q. W., Zhang, Z. Y., Yu, Y. S., Xiao, B., Wu, S. Y., Jiang, Z. L., Zhao, H. W., Zhao, R., and Li, J., Adenoviral vector mediates high expression levels of human lactoferrin in the milk of rabbits. J. Microbiol. Biotechnol., 18, 153–159 (2008).

    PubMed  CAS  Google Scholar 

  • Li, K., Li, Q., Li, J., Zhang, T., Han, Z., Gao, D., and Zheng, F., Antitumor activity of the procyanidins from Pinus koraiensis bark on mice bearing U14 cervical cancer. Yakugaku Zasshi, 127, 1145–1151 (2007).

    Article  PubMed  CAS  Google Scholar 

  • Mader, J. S., Salsman, J., Conrad, D. M., and Hoskin, D. W., Bovine lactoferricin selectively induces apoptosis in human leukemia and carcinoma cell lines. Mol. Cancer Ther., 4, 612–624 (2005).

    Article  PubMed  CAS  Google Scholar 

  • Mlejnek, P., Caspase inhibition and N6-benzyladenosineinduced apoptosis in HL-60 cells. J. Cell. Biochem., 83, 678–689 (2001).

    Article  PubMed  CAS  Google Scholar 

  • Mulder, A. M., Connellan, P. A., Oliver, C. J., Morris, C. A., and Stevenson, L. M., Bovine lactoferrin supplementation supports immune and antioxidant status in healthy human males. Nutr. Res., 28, 583–589 (2008).

    Article  PubMed  CAS  Google Scholar 

  • Ohtsuka, T., Buchsbaum, D., Oliver, P., Makhija, S., Kimberly, R., and Zhou, T., Synergistic induction of tumor cell apoptosis by death receptor antibody and chemotherapy agent through JNK/p38 and mitochondrial death pathway. Oncogene, 22, 2034–2044 (2003).

    Article  PubMed  CAS  Google Scholar 

  • Reed, J. C., Mechanisms of apoptosis. Am. J. Pathol., 157, 1415–1430 (2000).

    Article  PubMed  CAS  Google Scholar 

  • Sakai, T., Banno, Y., Kato, Y., Nozawa, Y., and Kawaguchi, M., Pepsin-digested bovine lactoferrin induces apoptotic cell death with JNK/SAPK activation in oral cancer cells. J. Pharmacol. Sci., 98, 41–48 (2005).

    Article  PubMed  CAS  Google Scholar 

  • Scorrano, L. and Korsmeyer, S. J., Mechanisms of cytochrome c release by proapoptotic BCL-2 family members. Biochem. Biophys. Res. Commun., 304, 437–444 (2003).

    Article  PubMed  CAS  Google Scholar 

  • Sekine, K., Watanabe, E., Nakamura, J., Takasuka, N., Kim, D. J., Asamoto, M., Krutovskikh, V., Baba-Toriyama, H., Ota, T., Moore, M. A., Masuda, M. A., Sugimoto, H., Nishino, H., Kakizoe, T., and Tsuda, H., Inhibition of azoxymethane-initiated colon tumor by bovine lactoferrin administration in F344 rats. Jpn. J. Cancer Res., 88, 523–526 (1997).

    PubMed  CAS  Google Scholar 

  • Shimazaki, K., Lactoferrin: A marvellous protein in milk? Anim. Sci. J., 71, 329–347 (2000).

    CAS  Google Scholar 

  • Sjöström, J., Blomqvist, C., von Boguslawski, K., Bengtsson, N. O, Mjaaland, I., Malmström, P., Ostenstadt, B., Wist, E., Valvere, V., Takayama, S., Reed, J. C., and Saksela, E., The predictive value of Bcl-2, Bax, Bcl-xL, bag-1, fas, and fasL for chemotherapy response in advanced breast cancer. Clin. Cancer Res., 8, 811–816 (2002).

    PubMed  Google Scholar 

  • Sun, G. P., Wang, H., Xu, S. P., Shen, Y. X., Wu, Q., Chen, Z. D., and Wei, W., Anti-tumor effects of paeonol in a HepAhepatoma bearing mouse model via induction of tumor cell apoptosis and stimulation of IL-2 and TNF-α production. Eur. J. Pharmacol., 584, 246–252 (2008).

    Article  PubMed  CAS  Google Scholar 

  • Takahashi, A., Masuda, A., Sun, M., Centonze, V. E., and Herman, B., Oxidative stress-induced apoptosis is associated with alterations in mitochondrial caspase activity and Bcl-2-dependent alterations in mitochondrial pH (pHm). Brain Res. Bull., 62, 497–504 (2004).

    Article  PubMed  CAS  Google Scholar 

  • Tsuda, H., Sekine, K., Fujita, K., and Ligo, M., Cancer prevention by bovine lactoferrin and underlying mechanisms -a review of experimental and clinical studies. Biochem. Cell Biol., 80, 131–136 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Tutykhina, I. L., Bezborodova, O. A., Shmarov, M. M., Logunov, D. Y., Neugodova, G. L., Nemtsova, E. R., Naroditsky, B. S., Yakubovskaya, R. I., and Gintsburg, A. L., Production of recombinant human lactoferrin in the allantoic fluid of embryonated chicken eggs and its characteristics. Protein Expr. Purif., 65, 100–107 (2009).

    Article  PubMed  CAS  Google Scholar 

  • Ward, P. P., Uribe-Luna, S., and Conneely, O. M., Lactoferrin and host defense. Biochem. Cell Biol., 80, 95–102 (2002).

    Article  PubMed  CAS  Google Scholar 

  • Ward, P. P. and Conneely, O. M., Lactoferrin: Role in iron homeostasis and host defense against microbial infection. Biometals, 17, 203–208 (2004).

    Article  PubMed  CAS  Google Scholar 

  • Wang, J., Li, Q., Ou, Y., Han, Z., and Li, K., Immunoregulation effect of recombinant adenoviral-mediated human lactoferrin gene on mice bearing EMT6 breast cancer. J. Pharm. Res., 3, 2131–2133 (2010).

    Google Scholar 

  • Xiao, Y., Monitto, C. L., Minhas, K. M., and Sidransky, D., Lactoferrin down-regulates G1 cyclin-dependent kinases during growth arrest of head and neck cancer cells. Clin. Cancer Res., 10, 8683–8686 (2004).

    Article  PubMed  CAS  Google Scholar 

  • Zhang, T., Li, Q., Li, K., Li, Y., Li, J., Wang, G., and Zhou, S., Antitumor effects of saponin extract from Patrinia villosa (Thunb.) Juss on mice bearing U14 cervical cancer. Phytother. Res., 22, 640–645 (2008).

    Article  PubMed  Google Scholar 

  • Zimecki, M., Artym, J., Chodaczek, G., Kocieba, M., Kuryszko, J., Houszka, M., and Kruzel, M. L., Immunoregulatory function of lactoferrin in immunosuppressed and autoimmune animals. Postepy Hig. Med. Dosw. (Online), 61, 283–287 (2007).

    Google Scholar 

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Correspondence to Qingwang Li.

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Wang, J., Li, Q., Ou, Y. et al. Inhibition of tumor growth by recombinant adenovirus containing human lactoferrin through inducing tumor cell apoptosis in mice bearing EMT6 breast cancer. Arch. Pharm. Res. 34, 987–995 (2011). https://doi.org/10.1007/s12272-011-0616-z

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  • DOI: https://doi.org/10.1007/s12272-011-0616-z

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