[In vitro and in vivo evaluation of the safety of Ad-RA538].
OBJECTIVE
To assess the safety of adenovirus-mediated transfer of the RA538 (Ad-RA538) for the treatment of cancer and to furthermore in preparation for a clinical trial of Ad-RA538.


METHODS
RT-PCR was used to detect the transcription of Ad-RA538 in HeLa cells infected with extracts from HeLa cells previously infected with Ad5-RA538. Cell counting was made to observe the effects of Ad-RA538 on the growth of the normal human fetal lung cell line 2BS. The virus was intraperitoneally injected into 2 groups of BalB/C mice at a dosage of 10(7) pfu and 10(9) pfu. Blood samples were taken from the mice to test the liver and renal function. PCR were used to screen the vital organs for the presence of adenovirus DNA. Microscopic examination of the vital organs was performed to observe the pathogenicity of Ad-RA538.


RESULTS
Ad-RA538 was a replication-defective virus. It could infect 2BS cells effectively, but could not inhibit 2BS cell growth. No mouse died and no signs of general toxicity were seen following intraperitoneal injection of Ad-RA538. The adenoviral vector was present in the liver, spleen, kidney and stomach of mice injected with 10(9) pfu Ad-RA538. Six and 12 days after injection, mild inflammation was observed in the liver of mice received 10(9) pfu Ad-RA538.


CONCLUSIONS
Ad-RA538 is safe both in vivo and in vitro, and clinical trials of Ad-RA538 can be performed.
Zhang
H
H
Department of Abdominal Surgery, Cancer Hospital, National Laboratory of Molecular Oncology, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, 100021.
Lin
C
C
Wei
Y
Y
D017498:Adenovirus E1 Proteins / Q000235:genetics*; D000260:Adenoviruses, Human / Q000235:genetics; D000818:Animals; D002471:Cell Transformation, Neoplastic / Q000187:drug effects; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004274:DNA, Recombinant / Q000096:biosynthesis / Q000633:toxicity*; D004279:DNA, Viral / Q000032:analysis; D004938:Esophageal Neoplasms / Q000473:pathology; D005822:Genetic Vectors; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D017498:Adenovirus E1 Proteins; D018076:DNA, Complementary; D004274:DNA, Recombinant; D004279:DNA, Viral; D014212:Tretinoin