Construction of PTEN eukaryotic expression plasmid and its effects on breast carcinoma cell line MDA468 / 中华肿瘤杂志

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of exogenous wild PTEN gene stably transfection on growth of breast cancer cells in vitro.</p><p><b>METHODS</b>At first, a recombinant eukaryotic expression plasmid pcDNA3.1-PTEN was constructed. Human breast cancer cell line MDA468 was transfected with pcDNA3.1-PTEN or mock transfected plasmid pcDNA3.1(-) with lipofectamine. RT-PCR, immunohistochemical staining and Western blot were used to determine target gene expression. Cell viability was tested by MTT assay. Apoptosis was determined by flow cytometry with a double-staining method using FITC-conjugated annexin V and PI.</p><p><b>RESULTS</b>The PTEN stably transfected cells demonstrated the integration of the exogenous target gene and corresponding mRNA and protein over-expression. There was a significant decline in cell viability of pcDNA3.1-PTEN transfected MDA468 cells in comparison with the mock-transfected ones (P < 0.01). The PTEN-trasfected MDA468 cells also showed an increase in the rate of apoptosis, compared with parental and mock-trasfected cells (P < 0.001).</p><p><b>CONCLUSION</b>Stable expression of exogenous PTEN can suppress the malignant phenotypes of the human breast cancer cell line MDA468.</p>