MiR-29a and miR-29c Recombinant Adenovirus Vector Regulate Proliferation of Human Bladder Cancer Cells

Abstract: To construct recombinant adenovirus vector containing human miR-29a and miR-29c and to determine its effect on the proliferation in human bladder cancer T24 cells， the PCR product containing miR-29a/c was amplified from human genomic DNA and inserted into the adenoviral shuttle vector pAdTrace-TO4-CMV. Then， the recombinant shuttle plasmid linearized by pme I was co-transformed into competent E. coli. BJ5183 with the adenoviral backbone plasmid pAdEasy-1. Then， the recombinant adenoviral DNA was transfected into HEK293 cells， packed and amplified miR-29a and miR-29c adenoviruses. T24 cells were infected by Ad-miR-29a and Ad-miR-29c. The expression of mature miR-29a/c was detected by Real-time PCR. The cell proliferation was detected by CCK-8 assay before and after infected miR-29a/c. Real-time PCR showed that miR-29a and miR-29c significantly up-regulated in T24 cells infected with Ad-miR-29a and Ad-miR-29c (P<0.01). CCK-8 assay showed that cell proliferation was significantly depressed after infection (P<0.05). The adenovirus vector Ad-miR-29a and Ad-miR-29c were constructed successfully and the over-expression of miR-29c inhibited the proliferation of T24 cells.

CSTR: 32200.14.cjcb.2014.02.0005