Influence of RNA Interference Targeting Rab5a on Proliferation and Invasion of Breast Cancer Cell Line MCF-7

ABSTRACT

Objective: To explore the influence of RNA interference targeting Rab5a on proliferation and invasion of breast cancer cells, MCF-7, in vitro and provide a new direction for breast cancer gene therapy. Method: Four miRNA interference vectors targeting Rab5a were designed, constructed, and transfected into breast cancer cell line MCF-7. RT-PCR and Western Blots were used to select the most efficient group of miRNA interference suppression. Next, the MTT and Transwell assays were applied to analyze cell proliferation inhibition rate and cell invasiveness.

Results: RT-PCR analysis showed that MCF-7 cells transfected with the Rab5a-miRNA-2 vector inhibited the Rab5a gene most strongly, with the inhibition rate being (50.01± 2.69)%. Western blot analysis also showed that cells transfected with Rab5a-miRNA-2 vector had the strongest inhibition of Rab5a protein, with the inhibition rate being (55.58± 3.16)%. Cells in this group were used Transwell assays. The Rab5a-miRNA-2 group was also used for MTT and Transwell assays. The MTT assay showed that the cell proliferation inhibitory rates were (2.47± 0.08)%, (19.69± 2.03)%, (24.81± 0.96)%, (32.90± 2.48)% at 12h, 36h, 60h, 84h since Rab5a-miRNA-2 vector were transfected into MCF-7, respectively. There was a significant difference between the blank and negative controls (P<0.01). The Transwell assay showed that the number of cells passing through the membranes was (67.8 ± 12.03) at 48h since transfection, with a significant difference between the negative controls and blank controls (P <0.01).

Conclusion: Rab5a can be silenced by miRNA expression vector thereby inhibiting the proliferation and invasiveness of MCF-7 breast cancer cells in vitro.