[The specific killing of human melanoma cells by replication selective adenovirus].
OBJECTIVE
To construct replication selective adenovirus AdhepE1 targeting human melanoma and observe its specific killing of human melanoma cells in vitro.


METHODS
Adenovirus E1 region, the murine tyrosinase promoter and enhancer DNA sequences were acquired respectively by PCR cloning. The shuttle plasmid of replication-selective adenovirus targeting human melanoma was constructed by DNA recombination. Replication-selective adenovirus AdhepE1 was generated by homologous recombination. The human melanoma cell line SK-Mel-1 and hepatocellular carcinoma cell line HepG2 were attacked separately by lower dose of AdhepE1. Change of cell morphology was observed and the surviving cells were calculated. The expression of E1A was assayed by RT-PCR to verify the specific-replication of AdhepE1.


RESULTS
Replication selective adenovirus AdhepE1 targeting human melanoma was acquired by PCR. Human melanoma cell line SK-Mel-1 was sensitive to oncolytic killing of AdhepE1 whereas HepG2 was little responsive. The results of RT-PCR suggested that AdhepE1 replicated specifically in human melanoma cells.


CONCLUSIONS
AdhepE1 can selectively kill human melanoma cells.
Xie
Qing-jun
QJ
Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
Lu
Ying-lin
YL
Chen
Ze-jian
ZJ
Zhang
Jin-qiang
JQ
Chen
Hui-hua
HH
Ling
Xian-long
XL
Lü
Pin
P
Du
Zhi-yan
ZY
Xu
Yuan-Ji
YJ
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D045744:Cell Line, Tumor; D015316:Genetic Therapy; D006801:Humans; D008113:Liver Neoplasms / Q000628:therapy; D008545:Melanoma / Q000628:therapy* / Q000821:virology; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014779:Virus Replication