[Anti-hepatocarcinoma effect of recombinant adenovirus carrying melittin gene].
OBJECTIVE
To find a new method of treating hepatocellular carcinoma with melittin by way of using the melittin gene.


METHODS
The recombinant adenoviruses carrying the melittin gene and alpha-fetoprotein (AFP) promoter (Ad-rAFP-Mel) were constructed through a bacterial homologous recombinant system. The efficiency of the adenovirus mediated gene transfer and the inhibition effect of Ad-rAFP-Mel on the proliferation of hepatocarcinoma cells were determined by X-gal staining and MTT assay respectively. The tumorigenicity of hepatocarcinoma cells transfected by Ad-rAFP-Mel and the antitumor effect of Ad-rAFP-Mel on the transplanted tumors in nude mice were detected in vivo.


RESULTS
The mRNA of the melittin gene was transcripted in HepG2 hepatocellular carcinoma cells transducted by Ad-rAFP-Mel. The efficiency of adenovirus mediated gene transfered to BEL-7402 hepatocarcinoma cells was 100% when the multiplicities of infection (MOI) of Ad-rAFP-Mel was 10 in vitro and was high in vivo as well. The inhibitive rates of Ad-rAFP-Mel and Ad-rAFP for BEL7402 cells were 66.2%+/-2.7% and 2.9%+/-2.3% (t = 30.83) by MTT assay. The inhibitive rates of Ad-CMV-Mel for BEL7402, SMMC7721 and L02 cells were 58.9%+/-9.6%, 65.9%+/-3.8%, 31.7%+/-1.2%, respectively, and those of the Ad-rAFP-Mel were 6.2%+/-2.7%, 16.1%+/-6.6%, 7.5%+/-3.3%, respectively (t = 1.27; t = 11.31, and t = 12.12, vs. Ad-CMV-Mel group in same cells). The tumorigenicity rates of hepatocarcinoma cells transfected by Ad-rAFP-Mel were decreased. A significant antineoplastic effect was detectd on transplanted tumor in nude mice by intratumoral injection of Ad-rAFP-Mel.


CONCLUSIONS
Ad-rAFP-Mel can inhibit specifically the proliferation of AFP-producing human hepatocarcinoma cells in vitro and in vivo. It suggests that animal toxin gene can be used as an interesting antitumor gene.
Ling
Chang-Quan
CQ
Department of Chinese Traditional Medicine, Changhai Hospital, Second Military Medical University, Shanghai 200433, China.
Li
Bai
B
Zhang
Chen
C
Gu
Wei
W
Li
Shao-Xiang
SX
Huang
Xue-Qiang
XQ
Zhang
Ya-Ni
YN
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D008114:Liver Neoplasms, Experimental / Q000473:pathology*; D008297:Male; D008555:Melitten / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects; D000509:alpha-Fetoproteins / Q000096:biosynthesis / Q000235:genetics
D011993:Recombinant Fusion Proteins; D000509:alpha-Fetoproteins; D008555:Melitten