Effects Of Suppressing MTA1 Expression Using RNA Interference Technique On Proliferation Of Human Breast Carcinoma Cell Line MDA-MB-231

Objective: The MTA1 gene has been recently identified as metastasis-associated gene and has been seen to correlate with the degree of invasion and metastasis in various carcinomas. The study aims to explore the effects of suppressing MTA1 expression using RNA interference technigue on the proliferation and cell cycle of human breast carcinoma cell line MDA-MB-231,and it could provide instructive value for the gene therapy of human breast carcinoma.Methods: A group of double strand oligonucleotide fragments were synthesized and cloned into pGenesil-1 vector. Lipofectamine 2000 transfection and G418 screening were used to establish MTA1 stable suppressed MDA-MB-231 cell lines. The MTA1 expression was assessed by RT-PCR;cell growth was evaluated by cell counting and MTT assay,and cell cycle was analyzed by flow cytometry. RT-PCR and immunohistochemistry were used to measure the expression of cyclinD1 in the MDA-MB-231 cells after suppressing of MTA1 .Results: Three MTA1 shRNAs have been designed and constructed into pGenesil-1 vector, and them were proved to be able to effectively depress MTA1 gene in MDA-MB-231 cells. The growth rate of MTA1 stably suppressed cells was significantly decreased and the cell cycle was stuck at G0～G1 phase.Conclusion: Suppression of the expression of MTA1 in MDA-MB-231 cells leads to cell proliferation inhibition and cell cycle arrest. RNAi-mediated inhibition of MTA1 may be a useful therapeutic approach for human breast carcinoma.