Biological Roles Of GSTM3 On Breast Cancer Cell Line MDA-MB-231

Objective:For further study on the role of GSTM3 in breast cancer, MDA-MB-231 breast cancer cell lines in which GSTM3 is overexpressed or silenced were constructed. The effects of overexpressing or silencing of GSTM3 on MDA-MB-231 cells were investigated.Methods:PCR was done to amplify the cDNA sequences of GSTM3 from prepared cDNA. The product was subcloned into pcDNA3.1 mammalian expression vector to obtain recombinant pcDNA3.1/GSTM3. The oligonucleotide sequences encoding the GSTM3 -specific siRNA were designed and subcloned into the vector pSilencer2.1-U6 neo, designated pSilencer-272, pSilencer-363 and pSilencer-529. These recombinant plasmids were transfected into MDA-MB-231 cells respectively. Expression level of GSTM3 mRNA and protein were analyzed by RT-PCR, real-time quantitative PCR and western blot. Using cell proliferation, cell migration assay, then the effect of GSTM3 on MDA-MB-231 cells proliferation and motility were investigated in vitro. Growth inhibition of ADM on cells was measured by the MTT assay.Results:1. Analyzed by semi-quantitative RT-PCR, real-time RT-PCR and Western blot, overexpression of GSTM3 was detected in cells transfected with pcDNA3.1/GSTM3.2. Analyzed by semi-quantitative RT-PCR and real-time RT-PCR, GSTM3 expression was significantly decreased in the siRNA interfered cell lines.3. The effects of GSTM3 overexpression or silenced on the breast cancer cell line MDA-MB-231:â‘ GSTM3 overexpression or silencing did not alter proliferation of MDA-MB-231 cells in vitro.â‘¡MDA-MB-231 cells motility was inhibited significantly by GSTM3 overexpression, and accelerated by GSTM3 silencing.â‘¢Overexpression of GSTM3 increased inhibitory effect of ADM on MDA-MB-231 cells, but the effect was reversed by GSTM3 silencing. Conclusion:1. MDA-MB-231 cell lines in which GSTM3 are stably overexpressed or silenced has been obtained.2. GSTM3 overexpression inhibited MDA-MB-231 cells motility, and enhanced inhibitory effect of ADM on MDA-MB-231 cells. And GSTM3 silencing promoted cells motility, and decreased inhibitory effect of ADM on MDA-MB-231 cells. The results indicated that GSTM3 might be associated with invasiveness and chemosensitivity of breast cancer cells.