Efficacy of recombinant adenovirus-mediated double suicide gene therapy in human keloid fibroblasts.
BACKGROUND
The recombinant adenovirus-mediated double suicide gene CDglyTK has been widely used for gene therapy of cancer. The biological behaviour of keloids is similar to that of cancer, in that they may extend beyond the site of injury, and do not subside. There are no effective strategies available for keloid therapy. Gene therapy is gaining greater importance in the field of plastic surgery, and the CDglyTK double suicide gene/prodrug system has been receiving greater attention.


OBJECTIVE
To show the lethal and bystander effects of a double suicide gene in keloid fibroblasts. Bcl-2 and BAX play an important role in the apoptosis induced by the double suicide gene.


METHODS
Recombinant adenovirus expression CDglyTK suicide genes were constructed using the modified AdEasy system. The lethal and bystander effects were measured after 48 h using the MTT assay. The morphological changes in fibroblasts were detected by haematoxylin and eosin staining, and apoptosis was detected by the terminal dUTP nick-end labelling assay. Bcl-2 and BAX were detected by immunohistochemistry and quantitative real-time PCR.


RESULTS
CDglyTK could be constructed easily and relatively quickly. The lethal and bystander effects of CDglyTK were marked in keloid fibroblasts. Apoptosis was one of the main processes leading to fibroblast death in keloids, and Bcl-2 and BAX played an important role in the process of apoptosis.


CONCLUSIONS
Together, the results support the notion that recombinant adenovirus-mediated CDglyTK double suicide gene therapy is effective in destroying keloid fibroblasts and provide a sound scientific rationale for keloid trials in vivo.
Xu
B
B
Department of Plastic Surgery, Second Hospital of Shandong University, Ji Nan, China.
Liu
Z Z
ZZ
Zhu
G Y
GY
Yang
J F
JF
Zhao
J P
JP
Wang
J C
JC
Cai
J L
JL
D000260:Adenoviruses, Human / Q000235:genetics*; D017209:Apoptosis / Q000235:genetics; D024201:Bystander Effect / Q000235:genetics*; D002460:Cell Line; D049109:Cell Proliferation; D004274:DNA, Recombinant / Q000235:genetics; D005347:Fibroblasts / Q000473:pathology; D041722:Genes, Transgenic, Suicide / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007627:Keloid / Q000473:pathology / Q000628:therapy*; D016896:Treatment Outcome
D004274:DNA, Recombinant