RESUMO
OBJECTIVE: To study the effects of mPer1 gene on the response of mammary carcinoma EMT6 cells to Adriamycin in vitro. METHODS: The eukaryotic expression vector pcDNA3. 1 (+)-mPer1 was transfected into the EMT6 cells (EMT6-mPerl). The vector pcDNA3. 1(+) transfect was also performed to serve as the control (EMT6-vect). The transfect efficiency was detected by RT-PCR and Western Blotting. The transfect cells were treated with Adriamycin in vitro. The apoptosis and distribution of cells in the cell cycle were analysed by FCM. The cell proliferation was detected by MTT assay. RT-PCR was used to show the mRNA expression of apoptosis-related genes. RESULTS: The mPerl-transfected EMT6 cells revealed S phase arrest, increased rate of apoptosis [EMT6-vect: (65.65 +/- 0.07)%; EMT6-mPer1: (72.35 +/- 0.57)%], decreased cell proliferation CEMT6-vect: (42.18 +/- 5.73)%; EMT6-mPer1: (53.28 +/- 7.32%)%] and stronger expression of p53 mRNA in RT-PCR (EMT6-vect, 0.48 +/- 0.08; EMT6-mPer1: 1.18 +/- 0.02). CONCLUSION: mPer1 gene can improve the drug sensitivity of this cell line to ADM in vitro.
