Construction of lentivirus-mediated shRNA vector targeting the human ITGβ1 gene and indentification of its effect on migration and invasion of tamoxifen-resistant MCF-7

Abstract:

Objective：To construct lentivirus vector targeting integrin β1（ITGβ1） gene and to study its effect on migration and inva－sion of Tamoxifen-resistant MCF-7（MCF-7R）. Methods：Four shRNA sequences targeting ITGβ1 gene and one negative control se－quence were designed，and were cloned into lentivirus vector，respectively. Recombinant lentivirus and control were extracted after transfecting 293T with the recombinant vector and helper vectors. The lentivirus with the best interfering effect was determined by real-time PCR and was chosen to infect MCF-7R. The expression of ITGβ1 gene was determined by Western blot. Transwell assay was used to detect the difference of migration and invasion. Results：Four recombinant lentivirus vectors were successfully constructed and the packaged lentivirus with the best interfering effect was used to infect MCF-7R. Western blot results showed that the protein expression of ITGβ1 was significantly reduced in MCF-7R（P=0.000） and its migration and invasion abilities were markedly suppressed（P=0.000）. Conclusion：Lentivirus vector targeting ITGβ1 gene can efficiently suppress the expression of ITGβ1 in MCF-7R，thereby leading to decreased migration and invasiveness abilities of MCF-7R