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[Pro-apoptotic effect on osteosarcoma SOSP-9607 cells by human recombinant caspase-6 fusion protein].

AbstractOBJECTIVE:
To investigate the pro-apoptotic effect of Her-2 targeted recombinant caspase-6 fusion protein on osteosarcoma SOSP-9607 cells.
METHODS:
Recombinant immunocasp-6 was generated by sequential fusion of the genes of a signal peptide, a single-chain Her-2 antibody (e23sFv), a PEA translocation domain (PEA aa253-364) and an active caspase-6. The immunocasp-6 gene was cloned into pCMV plasmid to construct a kind of eukaryotic expression vector, i.e. pCMV-e23sfv-PE II-caspase-6 (abbr. pCMV-6) and transfected into SOSP-9607 cells. Murine xenograft models were randomly divided into two groups that received i.m. injections of liposome encapsulated pCMV-6 or pCMV alone. The tumor volume and weight of the nude mice and the tumor weight of the cured mice were observed and statistically analyzed. The morphological changes of the tumors were examined with HE staining, apoptotic morphology of the tumor was observed by TUNEL staining and the gene expression was analyzed by immunohistochemical staining.
RESULTS:
The tumor growth of the mice in the treatment group was significantly slower than that of the control group (P = 0.001). The weight of the nude mice in the treatment group was significantly higher than that of the control group (P = 0.0002). The tumor weight of the mice in the treatment group was significantly lower than that of the control group (P = 0.0006). HE and TUNEL staining of the tumor of nude mice in the treatment groups showed typical characteristics of apoptosis, while normal structure was found in the control group. Furthermore, caspase-6 was not found in the tumor and muscle tissues in the control group, but only in the treatment group by immunohistochemistry.
CONCLUSION:
Immunocasp-6 can selectively recognize and bind to and kill HER-2 positive osteosarcoma cells, therefore, to offer some foundation for the clinical treatment of osteosarcoma.
AuthorsBen-gen Zhou, Xiu-chun Qiu, Yan-ming Xu, Qing-yu Fan
JournalZhonghua zhong liu za zhi [Chinese journal of oncology] (Zhonghua Zhong Liu Za Zhi) Vol. 32 Issue 7 Pg. 497-500 (Jul 2010) ISSN: 0253-3766 [Print] China
PMID21029691 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Bacterial Toxins
  • Exotoxins
  • Recombinant Fusion Proteins
  • Virulence Factors
  • immunocasp-6
  • ADP Ribose Transferases
  • Pseudomonas aeruginosa exotoxin A
  • Receptor, ErbB-2
  • Caspase 6
Topics
  • ADP Ribose Transferases (genetics)
  • Animals
  • Apoptosis
  • Bacterial Toxins (genetics)
  • Bone Neoplasms (metabolism, pathology)
  • Caspase 6 (genetics, metabolism)
  • Cell Line, Tumor
  • Exotoxins (genetics)
  • Humans
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • Neoplasm Transplantation
  • Osteosarcoma (metabolism, pathology)
  • Plasmids
  • Random Allocation
  • Receptor, ErbB-2 (genetics)
  • Recombinant Fusion Proteins (genetics, metabolism)
  • Transfection
  • Tumor Burden
  • Virulence Factors (genetics)

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