Effect of phosphatase activity of PTEN on migration ability of human breast cancer cell line ZR-75-1
- Author:
Guan-Ping LIN
1
Author Information
1. Department of Biochemistry and Molecular Biology
- Publication Type:Journal Article
- Keywords:
Breast neoplasms;
Human breast cancer cells ZR-75-1;
PTEN phosphohydrolase;
Transfectin
- From:
Tumor
2007;27(10):772-776
- CountryChina
- Language:Chinese
-
Abstract:
Objective: This study was to explore the effects of phosphatase activity of PTEN (phosphatase and tensin homolog deleted on chromosome 10) on migration ability of human breast cancer ZR-75-1 cells. Methods: Three kinds of plasmids, wt-PTEN plasmid, G129R-PTEN plasmid, and G129E-PTEN plasmid, were constructed and transfected into human breast cancer cell line ZR-75-1, with a deletion of PTEN gene, via liposome mediation. The expression of PTEN protein as well as the Tyr397 in phospho-focal adhesion kinase (p397.FAK) was measured by Western blotting. The effect of phosphatase activity of PTEN was observed in a cell scratch wound model in vitro. The inhibitory effects of phosphatase activity of PTEN were measured by using cell-matrix adhesion test and reconstituted basement membrane invasion technique. The expression level of MMP-2 was determined by immunohistochemical staining. Results: Three kinds of plasmids (wt-PTEN, G129R-PTEN, and G129E-PTEN) were successfully transfected into ZR-75-1 cells, respectively. All the three stably transfected cells had PTEN protein expression. Transfection of wt-PTEN and G129R-PTEN inhibited the migration of ZR-75-1 cells. There was significant difference in the inhibitory degree of cell adhesion and invasion between wt-PTEN or G129E-PTEN groups and G129R-PTEN transfected or non transfected groups (P < 0.01). The level of p397.FAK was significantly lower in wt-PTEN and G129E-PTEN groups compared with G129R-PTEN transfected groups. There was significant difference in the expression level of MMP-2 between G129R-PTEN-transfected and non transfected groups(P < 0.01). Conclusion: Both wild-type PTEN with dual specific phosphatase activities and G129E-PTEN with only protein phosphatase activity have inhibitory effects on migration of human breast cancer cell line ZR-75-1. However G129R-PTEN without phosphatase activity have no effects.
