Effect of CDK2-AP1 gene over-expression on proliferation and cell cycle regulation of breast cancer cell line MCF-7.
10.3969/j.issn.1672-7347.2012.10.004
- Author:
Xiaoyan GUAN
1
;
Weibing ZHOU
;
Juan HUANG
;
Longyun WANG
;
Yuping LIAO
Author Information
1. Department of Oncology, Central South University, Changsha, China.
- Publication Type:Journal Article
- MeSH:
Breast Neoplasms;
Cell Cycle;
Cell Division;
Cell Proliferation;
Cyclin-Dependent Kinases;
Down-Regulation;
Humans;
MCF-7 Cells;
Protein Kinases;
genetics;
metabolism;
Tumor Suppressor Proteins;
genetics;
metabolism
- From:
Journal of Central South University(Medical Sciences)
2012;37(10):990-996
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To over-express cyclin-dependent kinase 2-associated protein 1 (CDK2-AP1) gene, and investigate its effect on the proliferation and cell cycle regulation in breast cancer cell line MCF-7.
METHODS:CDK2-AP1 gene coding region was cloned into lentivirus vector. Lentivirus particles were infected into MCF-7 cells to upregulate the expression of CDK2-AP1 gene. The expression level of CDK2-AP1 was detected at both mRNA and protein levels by real-time PCR and Western blot. MTT assay, colony formatting assay, and flow cytometry were performed to detect the change of proliferation and cell cycle in MCF-7 cells. We examined the expression of cell cycle associated genes (CDK2, CDK4, P16Ink4A, and P21Cip1/Waf1) followed by CDK2-AP1 over-expression by Western blot.
RESULTS:CDK2-AP1 gene was up-regulated significantly at both mRNA (6.94 folds) and protein level. MTT based growth curve, colony formatting assay and flow cytometry showed that CDK2-AP1 over-expression lentivirus inhibited the proliferation of MCF-7 cells with statistical difference (P<0.05). In addition, with CDK2-AP1 over-expression, MCF-7 cells were arrested in G1 phase accompanied by apoptosis. Western blot showed that the expression level of P21Cip1/Waf1 and P16 Ink4A was upregulated, while the expression level of CDK2 and CDK4, members of the CDK family, was downregulated.
CONCLUSION:CDK2-AP1 gene plays a cancer suppressor role in breast cancer. Its function includes inhibiting the proliferation of MCF-7 cells and arresting the cell cycle in G1 phase.
