title: In Vitro Effects of HAS-2 Gene Silencing on the Proliferation and Apoptosis of the MCF-7 Human Breast Cancer Cell Line.
abstract: BACKGROUND
  Breast cancer is characterized by a distinct metastatic pattern involving the regional lymph nodes, bone marrow, lung and liver. This study is aimed to investigate the effects of silencing the HAS-2 gene on the proliferation and apoptosis of human breast cancer cells.
  
  
  METHODS
  MCF-7 cells were collected and assigned into control, scrambled siRNA and HAS-2- siRNA groups. After transfection, the morphological changes in the MCF-7 cells were observed using phase contrast microscopy. qRT-PCR and Western blot assays were used to detect the mRNA and protein expression of apoptosis-related proteins. CCK-8 and flow cytometry were performed to evaluate cell proliferation, the cell cycle and apoptosis.
  
  
  RESULTS
  In the control and the scrambled siRNA groups, cells grew adhered to the wall and mainly showed a spindle shape with a clear nucleolus. Compared with the control and scrambled siRNA groups, increases in the number of cells in early apoptosis and metaphase cells in apoptosis were observed in the HAS-2-siRNA group. The HAS-2-siRNA group showed decreased expression of HAS-2 relative to that in the control and scrambled siRNA groups. No significant differences in cell proliferation, cell cycle distribution or apoptosis were noted between the control and scrambled siRNA groups. In the HAS-2-siRNA group, the cell proliferation ability decreased significantly, but the number of cells in the G0/G1 stage, the number of apoptotic cells and the expression of caspase-3 and caspase-9 increased significantly.
  
  
  CONCLUSIONS
  Our findings indicate that HAS-2 gene silencing may inhibit proliferation and promote apoptosis in the MCF-7 human breast cancer cell line.
authors.affiliation: Department of Oncology, PLA Army General Hospital, Beijing, P.R. China.
authors.forename: Hong-Yan
authors.initials: HY
authors.lastname: Zhang
authors.forename: Feng
authors.initials: F
authors.lastname: Liang
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
authors.forename: Jian-Wei
authors.initials: JW
authors.lastname: Zhang
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Xi-Gang
authors.initials: XG
authors.lastname: Kang
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
authors.forename: Qi-Liu
authors.initials: QL
authors.lastname: Duan
mesh_terms: D017209:Apoptosis; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology* / Q000648:ultrastructure; D002453:Cell Cycle; D049109:Cell Proliferation; D048430:Cell Shape; D002999:Clone Cells; D005260:Female; D020868:Gene Silencing; D014453:Glucuronosyltransferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D000076002:Hyaluronan Synthases; D061986:MCF-7 Cells; D015398:Signal Transduction / Q000235:genetics; D014162:Transfection
chemical_list: D014453:Glucuronosyltransferase; C101754:HAS2 protein, human; D000076002:Hyaluronan Synthases