[Targeted tumor suppression by a secreted fusion protein consisting of anti- erbB2 antibody and reversed caspase-3 to SKBr3 cells].
OBJECTIVE
To investigate the targeted killing effect to SKBr3 cells due to the expression of a secreted fusion protein consisting of anti-erbB2 antibody and reversed caspase-3.


METHODS
A recombinant plasmid pCMV-e23scFv-PEII-revcasp 3 was constructed by subcloning reversed caspase-3 gene to the downstream of anti-erbB2 antibody and transfected into Jurkat cells. The cell lines which secreted expressing fusion protein stably were selected. The fusion protein in media was detected by ELISA and the media was used to culture human breast cancer SKBr3 cells. The recombinant plasmids with liposomes was administrated to BALB/C nude mouses bearing SKBr3 tumor by intramuscular injection. The targetting effect of the recombinant fusion protein caspase-3 was detected by indirect immunofluorescence staining.


RESULTS
Fusion protein can be expressed and secreted by Jurkat cells stably and kill SKBr3 cells. Significant prolonged survival time (prolonged by 72%) and inhibition of tumor growth in vivo (within inhibition ratio of 77%) were seen in the group administered with recombinant plasmids. Indirect immunofluorescence staining showed that the recombinant fusion protein caspase-3 has targetting effect.


CONCLUSIONS
Secreted expression of the fusion protein consisting of anti-erbB2 antibody and reversed caspase-3 can targetedly induce SKBr3 cells to death.
Zhang
Li-hong
LH
Department of Biochemistry and Molecular Biology, Faculty of Preclinical Medicine, Fourth Military Medical University, Xi'an 710032, China.
Jia
Lin-tao
LT
Yu
Cui-juan
CJ
Qu
Ping
P
Dong
Hai-long
HL
Zhao
Jing
J
Xu
Yan-ming
YM
Wang
Cheng-Ji
CJ
Yang
An-gang
AG
D000818:Animals; D000906:Antibodies / Q000235:genetics*; D053148:Caspase 3; D020169:Caspases / Q000235:genetics*; D015316:Genetic Therapy; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D009374:Neoplasms, Experimental / Q000628:therapy*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000037:antagonists & inhibitors*; D011993:Recombinant Fusion Proteins / Q000235:genetics*
D000906:Antibodies; D053148:Caspase 3; D020169:Caspases; D018719:Erb-b2 Receptor Tyrosine Kinases; D011993:Recombinant Fusion Proteins; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse