Cell Biochemistry and Biophysics

, Volume 70, Issue 3, pp 1893–1900 | Cite as

Anticancer Effects of Chemokine Receptor 4(CXCR4) Gene Silenced by CXCR4-siRNA in Nude Mice Model of Ovarian Cancer

Original Paper

Abstract

The aim is to study the anticancer effect of CXCR4 gene knockdown by CXCR4-siRNA in nude mice model of ovarian cancer. Injecti the SW626 tumor cells which had been transfected by vectors to make nude mouse model of ovarian cancer. The model mice were divided into interference group, negative control group, and blank control group. When the level of target genes were knocked down, the tumor volume was monitored and the tumor quality was measured; the expression of CXCR4 gene in the xenograft tumor was detected by RT-PCR, Western blot, and immunohistochemical staining. Nude mice model with implanted tumor were built successfully, after observing for 20 days. While the CXCR4 was knocked down, the abilities of invasion were weakened; the tumor volume and the tumor quality were also decreased. The CXCR4 mRNA and protein of the interference group decreased significantly (P < 0.05). The animal experiment was confirmed that silencing of CXCR4 gene by siRNA can obviously inhibit the tumorigenesis of ovarian cancer. Our work will provide the theoretical basis for genes interference therapy of ovarian cancer in future.

Keywords

Ovarian neoplasms Chemotactic factors Receptors CXCR4 RNA small interfering Nude mice Implanted tumor 

References

  1. 1.
    Zagouri, F., Dimopoulos, M. A., Bournakis, E., et al. (2010). Molecular markers in ovarian cancer: Their role in prognosis and therapy. European Journal of Gynaecological Oncology, 31(3), 268–277.PubMedGoogle Scholar
  2. 2.
    Ghezzi, F., Cromi, A., Siesto, G., et al. (2009). Laparoscopy staging of early ovarian cancer: Our experience and review of the literature. International Journal of Gynecological Cancer, 19(Suppl 2), S7–S13.PubMedCrossRefGoogle Scholar
  3. 3.
    Dinh, P., Harnett, P., Piccart-Gebhart, M. J., et al. (2008). New therapies for ovarian cancer: Cytotoxics and molecularly targeted agents. Critical Reviews in Oncology Hematology, 67(2), 103–112.CrossRefGoogle Scholar
  4. 4.
    Connolly, D. C., & Hensley, H. H. (2009). Xenograft and transgenic mice models of epithelial ovarian cancer and non invasive imaging modalities to monitor ovarian tumor growth in situ—applications in evaluating novel therapeutic agents. Current Protocols is Pharmacology, 45(14), 1211–1226.Google Scholar
  5. 5.
    Hai-ying, C., Jing-mao, W., Hong-ying, W., et al. (2012). Effect of short hairpin RNA-induced CXCR4 silence on ovarian cancer cell. Biomedicine & Pharmacotherapy, 66, 549–553.CrossRefGoogle Scholar
  6. 6.
    Itamochi, H., Kigawa, J., Kanamori, Y., et al. (2007). Adenovirus type 5 E1A gene therapy for ovarian clear cell carcinoma: A potential treatment strategy. Molecular Cancer Therapeutics, 6(1), 227–235.PubMedCrossRefGoogle Scholar
  7. 7.
    Elbashir, S. M., Harborth, J., Lendeckel, W., et al. (2001). Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature, 411(6836), 494–498.PubMedCrossRefGoogle Scholar
  8. 8.
    Chen J., Yan, L. B., Officer Spring Cloud. (2008). RNA interference and its application research. Journal of Hunan Agricultural University Agronomy Courtyard, 3(S1): 300–304.Google Scholar
  9. 9.
    Nicolas, F. E., Lopez-Gomollon, S., Lopez-Martinez, A. F., et al. (2009). RNA silencing: Recent developments on miRNAs. Recent Patents on DNA Gene Sequences, 3(2), 77–87.PubMedCrossRefGoogle Scholar
  10. 10.
    Couzin, J. (2002). Breakthrough of the year. Small RNAs make big splash. Science, 298(5602), 2296–2297.PubMedCrossRefGoogle Scholar
  11. 11.
    Salvi, A., Arici, B., De Petro, G., et al. (2004). Small interfering RNA urokinase small interfering RNA urokinase silencing inhibits invasion and migration of human hepatocellular carcinoma cells. Molecular Cancer Therapeutics, 3(6), 671–678.PubMedGoogle Scholar
  12. 12.
    Pardridge, W. M. (2004). Intravenous, non-viral RNAi gene therapy of brain cancer. Expert Opinion on Biological Therapy, 4(7), 1103–1113.PubMedCrossRefGoogle Scholar
  13. 13.
    Zhang, L., Yang, N., Mohamed-Hadley, A., et al. (2003). Vector-based RNAi, a novel tool for isoform-specific knock-down of VEGF and anti-angiogenesis gene therapy of cancer. Biochemical and Biophysical Research Communications, 303(4), 1169–1178.PubMedCrossRefGoogle Scholar
  14. 14.
    Liu, Y., Zhang, R., et al. (2007) CXCR4, its ehrs-2 expression in epithelial ovarian cancer and its correlation study. Journal of Modern Cancer Medicine, 15(11): 1669–1672.Google Scholar
  15. 15.
    Bajetto, A., Barbero, S., Bonavia, R., et al. (2001). Stromal cell-derived factor-1 alpha induces astrocyte proliferation through the activation of extracellular signal-regulated kinases 1/2 pathway. Journal of Neurochemistry, 77(5), 1226–1236.PubMedCrossRefGoogle Scholar
  16. 16.
    Liu, Z., & Habener, J. F. (2009). Stromal cell-derived factor-1 promotes survival of pancreatic beta cells by the stabilisation of beta-catenin and activation of transcription factor 7-like 2 (TCF7L2). Diabetologia, 52(8), 1589–1598.PubMedCentralPubMedCrossRefGoogle Scholar
  17. 17.
    Liang, Z., Yoon, Y., Votaw, J., et al. (2005). Silencing of CXCR4 blocks breast cancer metastasis. Cancer Research, 65(3), 967–971.PubMedCentralPubMedGoogle Scholar
  18. 18.
    Scala, S., Ottaiano, A., Ascierto, P. A., et al. (2005). Expression of CXCR4 predicts poor prognosis in patients with malignant melanoma. Clinical Cancer Research, 11(5), 1835–1841.PubMedCrossRefGoogle Scholar
  19. 19.
    Dziembowska, M., Tham, T. N., Lau, P., et al. (2005). A role for CXCR4 signaling in survival and migration of neural and oligodendrocyte precursors. Glia, 50(3), 258–269.PubMedCrossRefGoogle Scholar
  20. 20.
    Nimmagadda, S., Pullambhatla, M., & Pomper, M. G. (2009). Immuno imaging of CXCR4 expression in brain tumor xenografts using SPECT/CT. Journal of Nuclear Medicine, 50(7), 1124–1130.PubMedCentralPubMedCrossRefGoogle Scholar
  21. 21.
    Jaszczynska-Nowinka, K., & Markowska, A. (2009). New cytokine: Stromal derived factor-1. European Journal of Gynaecological Oncology, 30(2), 124–127.PubMedGoogle Scholar
  22. 22.
    Darash-Yahana, M., Pikarsky, E., Abramovitch, R., et al. (2004). Role of high expression levels of CXCR4 in tumor growth, vascularization, and metastasis. The FASEB Journal, 18(11), 1240–1242.Google Scholar
  23. 23.
    Shen, X. Y., Wang, S. H., Liang, M. L., et al. (2008). The role and mechanism of CXCR4 and its ligand SDF-1 in the development of cervical cancer metastasis. Ai Zheng, 27(10), 1044–1049.PubMedGoogle Scholar
  24. 24.
    Song, J. S., Kang, C. M., Kang, H. H., et al. (2010). Inhibitory effect of CXC chemokine receptor 4 antagonist AMD3100 on bleomycin induced murine pulmonary fibrosis. Experimental & Molecular Medicine, 42(6), 465–472.CrossRefGoogle Scholar
  25. 25.
    Barbieri, F., & Bajetto, A. (2010). Role of chemokine network in the development and progression of ovarian cancer: A potential novel pharmacological target. Journal of Oncology, 1(2010), 426956.Google Scholar
  26. 26.
    Wang, J., Cai, J., Han, F., et al. (2011). Silencing of CXCR4 blocks progression of ovarian cancer and depresses canonical Wnt signaling pathway. International Journal of Gynecological Cancer, 21(6), 981–987.PubMedCrossRefGoogle Scholar
  27. 27.
    Filleur, S., Courtin, A., Ait-Si-Ali, S., et al. (2003). SiRNA mediated inhibition of vascular endothelial growth factor severely limits tumor resistance to antiangiogenic thrombospondin-1 and slows tumor vascularization and growth. Cancer Research, 63, 3919–3922.PubMedGoogle Scholar
  28. 28.
    Yu, N. X., Zhilan, P., Weijiang, D., et al. (2006). RNA interference in cervical cancer HPV16E6 gene research. Journal of Sichuan University (Medical Edition), 37(1), 14–18.Google Scholar
  29. 29.
    Gomes-da-Silva, L. C., Fonseca, N. A., Moura, V., et al. (2012). Lipid-based nanoparticles for siRNA delivery in cancer therapy: Paradigms and challenges [J]. Accounts of Chemical Research, 45(7), 1163.PubMedCrossRefGoogle Scholar
  30. 30.
    Miyagishi, M., & Taira, K. (2002). Development and application of siRNA expression vector. Nucleic Acids Research, 2(suppl), 113–114.PubMedCrossRefGoogle Scholar
  31. 31.
    Dhiraj, B. K. S., Swam, S., & Uma, M. (2012). Translational siRNA therapeutics using liposomal carriers: Prospects & challenges. Current Gene Therapy, 12(18), 315.Google Scholar
  32. 32.
    Audouy, S. A., Leij, L. E., Hoekstra, D., & Molema, G. (2002). In vivo characteristics of cationic liposomes as delivery vectors for gene therapy. Pharmaceutical Research, 19(11), 1599–1605.PubMedCrossRefGoogle Scholar
  33. 33.
    Tousignant, J. D., Gates, A. I., lngram, L. A., et al. (2000). Comprehensive analysis of the acute toxicities induced by systemic administration of cationic lipid: Plasmid DNA complexes in mice. Human Gene Therapy, 11(18), 2493–2513.PubMedCrossRefGoogle Scholar
  34. 34.
    Schiffelers, R M Xu J, Stom, G., et al. (2005). Effects of treatment with small interfering RNA on joint inflammation in mice with collagen-induced arthritis. Arthritis and Rheumatism, 52(4), 1314–1318.PubMedCrossRefGoogle Scholar
  35. 35.
    Guoping, H., Sizhong, Z., Yingcheng, W., et al. (2005). Short hairpin RNA mediated time and dose effect of RNA interference study. Progress in Biochemistry and Biophysics, 32(3), 258–267.Google Scholar
  36. 36.
    Paddison, P. J., Caudy, A. A., Bernstein, E., et al. (2002). Short hairpin RNAs(shRNAs) induce sequence-specific silencing in mammalian cells. Genes & Development, 16(8), 948–958.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Wei Liu
    • 1
  • Yueyun Wang
    • 1
  • Hongying Wang
    • 1
  • Aixia Wang
    • 1
  1. 1.The Department of Obstetrics and GynecologyLiaocheng People’s HospitalLiaochengChina

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