title: [Effect of CDK2-AP1 gene over-expression on proliferation and cell cycle regulation of breast cancer cell line MCF-7].
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Oncology, Central South University, Changsha, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Guan
authors.forename: Weibing
authors.initials: W
authors.lastname: Zhou
authors.forename: Juan
authors.initials: J
authors.lastname: Huang
authors.forename: Longyun
authors.initials: L
authors.lastname: Wang
authors.forename: Yuping
authors.initials: Y
authors.lastname: Liao
mesh_terms: D001943:Breast Neoplasms; D002453:Cell Cycle; D002455:Cell Division; D049109:Cell Proliferation; D018844:Cyclin-Dependent Kinases; D015536:Down-Regulation; D006801:Humans; D061986:MCF-7 Cells; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D025521:Tumor Suppressor Proteins; D011494:Protein Kinases; D018844:Cyclin-Dependent Kinases