Construction of PTEN eukaryotic expression plasmid and its effects on breast carcinoma cell line MDA468.
- Author:
Qing-yong CHEN
1
;
Dao-da CHEN
;
Chun-you WANG
;
You-sheng ZHOU
Author Information
- Publication Type:Journal Article
- MeSH: Breast Neoplasms; metabolism; pathology; Cell Line, Tumor; Eukaryotic Cells; metabolism; Humans; PTEN Phosphohydrolase; biosynthesis; genetics; Phenotype; Plasmids; genetics; Recombinant Proteins; biosynthesis; genetics; Transfection
- From: Chinese Journal of Oncology 2005;27(4):216-219
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effects of exogenous wild PTEN gene stably transfection on growth of breast cancer cells in vitro.
METHODSAt 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.
RESULTSThe 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).
CONCLUSIONStable expression of exogenous PTEN can suppress the malignant phenotypes of the human breast cancer cell line MDA468.
