Effects of rAAV-CD151 and rAAV-antiCD151 on the migration of human tongue squamous carcinoma cell line Tca8113

Abstract

This study was designed to determine the effects of the recombinant adeno-associated virus vector containing sense CD151 gene (rAAV-CD151) and antisense CD151 gene (rAAV-antiCD151) on the migration of Tca8113 cell. Functional fragment of CD151 gene was amplified by RT-PCR, and inserted into the vector pAAV in the sense direction and antisense direction, respectively. The rAAV-CD151 and rAAV-antiCD151 were produced and the titers were determined by dot blot. The CD151, at protein level, was detected by Western blot. The Transwell chamber was used to detect the effects of the rAAV-CD151 and rAAV-antiCD151 on the tumor cell migration. The titers of the rAAV-CD151 and rAAV-antiCD151 were 2×10¹¹ pfu/ml and 1.0×10¹¹ pfu/ml, respectively. The expression of CD151 was increased by 108% in the cells transfected with rAAV-CD151 and decreased by 79% in the cells transfected with rAAV-antiCD151, as compared with non-transfected cells, respectively. The number of the migrating cells was significantly increased in the cells transfected with rAAV-CD151 (93.56±11.59) and decreased in the cells transfected with rAAV-antiCD151 (24.00±4.36) as compared with non-transfected and rAAV-GFP transfected cells (53.00±6.56 and 46.00±7.00,P<0.05). It is an important molecular mechanism of the tumor metastasis that the overexpression of CD151 promotes the migration of the tumor cells. The rAAV-antiCD151 is a novel tool, which can reduce the expression of CD151 and inhibit the migration of the tumor cells, and brings us a new approach of anti-sene gene therapy targeted at CD151 in human carcinoma.

Keywords

CD151 gene / rAAV / anti-sense nuclei acid / cell migration

References

[1] KazarovA R, YangX, StippC, et al. . An extracellular site on tetraspanin CD151 determines alpha 3 and alpha 6 integrin-dependent cellular morphology. J Cell Biol, 2002, 158: 1299-1299

[2] HemlerM E. Specific tetraspanin functionset al. J Cell Biol, 2001, 155: 1103-1103

[3] TokuharaT, HasegawaH, HattoriN, et al. . Clinical significance of CD151 gene expression in non-small cell lung cancer. Clin Cancer Res, 2001, 7: 4109-4109

[4] HashidaH, TakabayashiA, TokuharaT, et al. . Clinical significance of transmembrane 4 superfamily in colon cancer. Br J Cancer, 2003, 89: 158-158

[5] BerditchevskiF, GilbertE, GriffithsM R, et al. . Analysis of the CD151-alpha3betal integrin and CD151-tetraspanin interactions by mutagenesis. J Biol Chem, 2001, 276: 11165-11165

[6] ZhangX A, BomtragerA L, HemlerM E, et al. . Transmembrane-4 superfamily proteins associate with activated protein kinase C (PKC) and link PKC to specific beta(1) integrins. J Biol Chem, 2001, 276: 25005-25005

[7] BerditchevskiF. Complexes of tetraspanins with integrins: more than meets the eye. J Cell Sci, 2001, 111: 1113-1113

[8] MonahanP E, SamulskiR J, et al. . AAV: is clinical success in the horizon?. Gene Ther, 2000, 7: 21-21