[Apoptosis of human melanoma cell line WM-983A by p16 gene transduction].
OBJECTIVE
To further understand the mechanism of action of the tumor-suppressor gene p16.


METHODS
An adenoviral expression vector with full length cDNA of p16 gene insert was constructed (Ad-p16) and transfected into WM-983A cells, the p16 gene of which was point mutated at codon 126. The effect of exogenous p16 gene on the growth of WM-983A cells was examined in vitro and in vivo.


RESULTS
Expression of p16 gene in WM-983A cells was confirmed by Western blot. The in vitro growth of the Ad-p16 transfected WM-983A cells was significantly inhibited (inhibition rate: 78%) as compared to mock (Ad-LacZ) transfected WM-983A cells. Colony-forming activity in vitro of the Ad-p16 transfected WM-983A cells was completely inhibited. Morphologically, the Ad-p16 transfected cells appeared apoptotic which was confirmed by the appearance of pre-G1 on flow cytometry and DNA fragmentation. The growth of WM-983A xenografts in nude mice was retarded by intra-tumoral injection of Ad-p16.


CONCLUSIONS
p16 gene participates in the induction of cell apoptosis. It is promising to use it for gene therapy of cancer, especially when combined with other apopptosis-inducing agents.
Cheng
J
J
National Laboratory of Molecular Oncology, Cancer Institute, Chinese Academy of Medical Sciences (Hospital), Peking Union Medical College, Beijing 100021.
Lin
C
C
Xing
R
R
D000818:Animals; D017209:Apoptosis; D019942:Genes, p16; D015316:Genetic Therapy; D006801:Humans; D008545:Melanoma / Q000473:pathology / Q000628:therapy*; D051379:Mice; D008819:Mice, Nude; D014162:Transfection; D014407:Tumor Cells, Cultured