Expression and anticancer activity analysis of recombinant human uPA1-43-melittin
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..
| Medienart: |
Artikel |
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| Erscheinungsjahr: |
2015 |
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| Erschienen: |
2015 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:46 |
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| Enthalten in: |
International journal of oncology - 46(2015), 2, Seite 619-626 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
SU, MANMAN [VerfasserIn] |
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| Links: |
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| doi: |
10.3892/ijo.2014.2750 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
OLC1959538829 |
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| 100 | 1 | |a SU, MANMAN |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Expression and anticancer activity analysis of recombinant human uPA1-43-melittin |
| 264 | 1 | |c 2015 | |
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| 520 | |a 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. | ||
| 540 | |a Nutzungsrecht: Copyright © 2015, Spandidos Publications | ||
| 650 | 4 | |a melittin | |
| 650 | 4 | |a urokinase plasminogen activator | |
| 650 | 4 | |a Pichia pastoris | |
| 650 | 4 | |a ovarian cancer | |
| 650 | 4 | |a fusion protein | |
| 650 | 4 | |a Melitten - administration & dosage | |
| 650 | 4 | |a Ovarian Neoplasms - pathology | |
| 650 | 4 | |a Bee Venoms - genetics | |
| 650 | 4 | |a Urokinase-Type Plasminogen Activator - administration & dosage | |
| 650 | 4 | |a Recombinant Fusion Proteins - genetics | |
| 650 | 4 | |a Urokinase-Type Plasminogen Activator - genetics | |
| 650 | 4 | |a Bee Venoms - chemistry | |
| 650 | 4 | |a Ovarian Neoplasms - therapy | |
| 650 | 4 | |a Melitten - genetics | |
| 650 | 4 | |a Recombinant Fusion Proteins - administration & dosage | |
| 650 | 4 | |a Receptors, Urokinase Plasminogen Activator - genetics | |
| 650 | 4 | |a Ovarian Neoplasms - genetics | |
| 650 | 4 | |a Apoptosis - drug effects | |
| 650 | 4 | |a Gene Expression Regulation, Neoplastic - drug effects | |
| 650 | 4 | |a Cell Cycle Checkpoints - drug effects | |
| 700 | 1 | |a CHANG, WEIQIN |4 oth | |
| 700 | 1 | |a CUI, MANHUA |4 oth | |
| 700 | 1 | |a LIN, YANG |4 oth | |
| 700 | 1 | |a WU, SHUYING |4 oth | |
| 700 | 1 | |a XU, TIANMIN |4 oth | |
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| 773 | 1 | 8 | |g volume:46 |g year:2015 |g number:2 |g pages:619-626 |
| 856 | 4 | 1 | |u http://dx.doi.org/10.3892/ijo.2014.2750 |3 Volltext |
| 856 | 4 | 2 | |u http://www.ncbi.nlm.nih.gov/pubmed/25394558 |
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