Inhibitory effect of Bcl-2 short hairpin RNA on growth of BEL-7402 and Caco2 cells / 第二军医大学学报

ABSTRACT

Objective: To investigate the effects of Bcl-2 shRNA on the growth of liver carcinoma cell line BEL-7402 and colorectal carcinoma cell line Caco2. Methods: Bcl-2 shRNA was cloned into Pgenesil-1 plasmid labeled with fluorescent protein and the product was transfected into BEL-7402 and Caco2 cells with Lipofectamine 2000. The study also included shRNA as negative control, Pgenesil-1, Lipofectamine and blank control groups. Transfected cells were visualized by inverted fluorescent microscope and then assayed by flow cytometry. The expression levels of Bcl-2 protein were assayed by Western-blot and cell proliferation was measured by MTT method. Results: There was no difference in transfection rate among cells in Bcl-2 shRNA, shRNA and Pgenesil-1 vector groups. Western-blot assay showed that the expression levels of Bcl-2 protein in BEL-7402 and Caco2 cells were significantly decreased in Bcl-2 shRNA group compared with those in other 4 groups (P<0.05). Furthermore, Bcl-2 shRNA had a stronger inhibitory effect on Caco2 cells than on BEL-7402 cells (P<0.05). MTT assay showed that Bcl-2 shRNA significantly inhibited the growth of BEL-7402 and Caco2 cells (at 72 and 96 h after treatment, respectively) compared with the other 4 groups (P<0.05). Conclusion: Bcl-2 shRNA can specifically inhibit the growth of BEL-7402 and Caco2 cells.