Suppression of the tumorigenicity of B-16V melanoma cells via lysozyme gene.
OBJECTIVE
The aim of this study was to evaluate the effects of lysozyme on the tumorigenicity of B-16V melanoma cells.


METHODS
After performing a series of molecular biology applications, including mRNA isolation, reverse transcriptase polymerase chain reaction, restriction digestions and ligations, recombinant pHM6 vector harboring mouse lysozyme gene (pHM6mLys) was constructed. B-16V melanoma cells were transfected with plasmid DNAs (pHM6 and pHM6mLys). Transfected cells (B-16VpHM6 and B-16VpHM6mLys) were selected in media containing geneticin. B-16V, B-16VpHM6, and B-16VpHM6mLys cells were then injected subcutaneously (s.c.) to the three groups of C57BL/6 inbred mice (30 mice/group). These mice were examined every 3 days for s.c. tumor development over 41 days. The results were evaluated by using statistical methods.


RESULTS
Tumor formation was observed in all mice injected with B-16V and B-16VpHM6 cells in the first 8-12 days. However, tumor didn't develop in 16 of 30 of the mice injected with B-16VpHMmLys cells. Tumor-free animals (16 mice) in this group were reinjected with B-16V cells, and 9 of them died during the first 10 days of observation. Tumor development was not observed in the remaining 7 mice over 60 days of the experimental period. Results were statistically significant (p values < or = 0.05).


CONCLUSIONS
These findings indicate that lysozyme expressed by B-16VpHMmLys cells may suppress the tumorigenicity of these cells and may help development of protective immunity against B-16V melanoma cells.
Guzel
Ali I
AI
Department of Medical Biology and Genetics, Faculty of Medicine Cukurova University, Adana, Turkey.
Kasap
Halil
H
Tuncer
Ilhan
I
Ozgunen
Kerem
K
Sertdemir
Yasar
Y
Kasap
Mulkiye
M
Ozcan
Numan
N
Yilmaz
Mehmet B
MB
Pazarbasi
Ayfer
A
D000818:Animals; D003001:Cloning, Molecular; D015972:Gene Expression Regulation, Neoplastic; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D005839:Gentamicins / Q000494:pharmacology; D008545:Melanoma / Q000628:therapy*; D008546:Melanoma, Experimental / Q000628:therapy*; D051379:Mice; D015233:Models, Statistical; D009113:Muramidase / Q000235:genetics*; D016017:Odds Ratio; D010957:Plasmids / Q000378:metabolism; D013997:Time Factors; D016896:Treatment Outcome
D005839:Gentamicins; C010680:antibiotic G 418; D009113:Muramidase