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International Journal of Oncology
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International Journal of Molecular Medicine

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International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease.

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International Journal of Oncology is an international journal devoted to oncology research and cancer treatment.

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Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine.

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Article

Expression and anticancer activity analysis of recombinant human uPA1‑43-melittin

  • Authors:
    • Manman Su
    • Weiqin Chang
    • Manhua Cui
    • Yang Lin
    • Shuying Wu
    • Tianmin Xu
  • View Affiliations / Copyright

    Affiliations: The Second Hospital, Jilin University, Changchun, P.R. China
  • Pages: 619-626
    |
    Published online on: November 13, 2014
       https://doi.org/10.3892/ijo.2014.2750
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Abstract

The present study is focused on expression of a target fusion protein which can be used in ovarian cancer target therapy. It aimed to construct human urokinase-type plasminogen activator (uPA)1‑43-melittin eukaryotic expression vector to express recombinant human uPA1‑43-melittin (rhuPA1‑43-melittin) in P. pastoris and to detect its anticancer effects on ovarian cancer cells. The DNA sequences that encode uPA1‑43 amino acids and melittin were synthesized according to its native amino acid sequences and consequently inserted into pPICZαC vector. Then uPA1‑43-melittin -pPICZαC was transformed into P. pastoris X-33, and rhuPA1‑43-melittin was expressed by methonal inducing. The bioactivities of recombinant fusion protein were detected with inhibition effects on growth of ovarian cancer cells, cell cycle detection and TUNEL assay. The results of DNA sequence analysis of the recombinant vector uPA1‑43-melittin -pPICZαC demonstrated that the DNA encoding human uPA 1‑43 amino acids and 1-26 amino acids of melittin was correctly inserted into the pPICZαC vector. After being induced by methonal, fusion protein with molecular weight 7.6 kDa was observed on the basis of SDS-PAGE and western blot analysis. The recombinant protein was able to suppress growth of SKOV3, induce cell cycle arrest and apoptosis of SKOV3 cells. The fusion protein does not have any obvious toxicity on normal tissues. RhuPA1‑43-melittin was successfully expressed in P. pastoris. Taking uPA1‑43 amino acids specifically binding to uPAR as targeted part of fusion protein, and making use of antitumor activity of melittin, the recombinant fusion protein it was able to inhibit growth of ovarian tumors and to be applied for effective targeted treatment.
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Copy and paste a formatted citation
Spandidos Publications style
Su M, Chang W, Cui M, Lin Y, Wu S and Xu T: Expression and anticancer activity analysis of recombinant human uPA1‑43-melittin. Int J Oncol 46: 619-626, 2015.
APA
Su, M., Chang, W., Cui, M., Lin, Y., Wu, S., & Xu, T. (2015). Expression and anticancer activity analysis of recombinant human uPA1‑43-melittin. International Journal of Oncology, 46, 619-626. https://doi.org/10.3892/ijo.2014.2750
MLA
Su, M., Chang, W., Cui, M., Lin, Y., Wu, S., Xu, T."Expression and anticancer activity analysis of recombinant human uPA1‑43-melittin". International Journal of Oncology 46.2 (2015): 619-626.
Chicago
Su, M., Chang, W., Cui, M., Lin, Y., Wu, S., Xu, T."Expression and anticancer activity analysis of recombinant human uPA1‑43-melittin". International Journal of Oncology 46, no. 2 (2015): 619-626. https://doi.org/10.3892/ijo.2014.2750
Copy and paste a formatted citation
x
Spandidos Publications style
Su M, Chang W, Cui M, Lin Y, Wu S and Xu T: Expression and anticancer activity analysis of recombinant human uPA1‑43-melittin. Int J Oncol 46: 619-626, 2015.
APA
Su, M., Chang, W., Cui, M., Lin, Y., Wu, S., & Xu, T. (2015). Expression and anticancer activity analysis of recombinant human uPA1‑43-melittin. International Journal of Oncology, 46, 619-626. https://doi.org/10.3892/ijo.2014.2750
MLA
Su, M., Chang, W., Cui, M., Lin, Y., Wu, S., Xu, T."Expression and anticancer activity analysis of recombinant human uPA1‑43-melittin". International Journal of Oncology 46.2 (2015): 619-626.
Chicago
Su, M., Chang, W., Cui, M., Lin, Y., Wu, S., Xu, T."Expression and anticancer activity analysis of recombinant human uPA1‑43-melittin". International Journal of Oncology 46, no. 2 (2015): 619-626. https://doi.org/10.3892/ijo.2014.2750
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