Silencing Bmi-1 enhances the senescence and decreases the metastasis of human gastric cancer cells.

ABSTRACT:

Aim

To evaluate the impact of Bmi-1 on cell senescence and metastasis of human gastric cancer cell line BGC823.

Methods

Two pairs of complementary small hairpin RNA (shRNA) oligonucleotides targeting the Bmi-1 gene were designed, synthesized, annealed and cloned into the pRNAT-U6.2 vector. After DNA sequencing to verify the correct insertion of the shRNA sequences, the recombinant plasmids were transfected into BGC823 cells. The expression of Bmi-1 mRNA and protein was examined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. The effects of Bmi-1 knockdown on cell senescence and metastasis were determined by the β-Gal activity assay and Boyden chamber assay, respectively.

Results

The double-stranded oligonucleotide fragments of Bmi-1 s

SUBMITTER: Gao FL 

PROVIDER: S-EPMC3870526 | biostudies-literature | 2013 Dec

REPOSITORIES: biostudies-literature