title: [Effects of down-regulation of CC chemokine ligand 5 (CCL5) on proliferation of human breast cancer cells in vitro].
abstract: OBJECTIVE
  To investigate the effect of suppression of CCL5 ligand gene on the proliferation of human breast cancer cells.
  
  
  METHODS
  A lentiviral vector carrying a short interfering RNA (siRNA) targeting CCL5 was transfected into human breast cancer cell line MCF-7 and MDA-MB-231 cells. The expression of CCL5 mRNA in the cells was detected by real-time PCR. The proliferation of MCF-7 and MDA-MB-231 cells was assessed by MTT assay and FACS assay, and the colony formation ability of both cell lines were measured, respectively.
  
  
  RESULTS
  Real time PCR showed a good knockdown effect of CCL5 in both cell-lines. Colony-forming assay showed that the ability of colony formation of MCF-7/CCL5-siRNA and MDA-MB-231/CCL5-siRNA was decreased markedly. The colony number of MCF-7/CCL5-siRNA group was (0.34 ± 0.08), significantly lower than 0.81 ± 0.12 in the MCF-7/CCL5-N group and 0.92 ± 0.12 in the MCF-7 group (P < 0.05). The colony number of MDA-MB-231/CCL5-siRNA group was 0.33 ± 0.10, significantly lower than 0.97 ± 0.09 in the MDA-MB-231/CCL5-N group and 1.04 ± 0.07 in the MDA-MB-231 group (P < 0.05). However, MTT assay revealed that the proliferation of MCF-7/CCL5-siRNA cells was not significantly different from that of MCF-7/CCL5-N or MCF-7 cells, respectively (P > 0.05), and the same result was found in MDA-MB-231 cells. FACS assay showed that the proliferation index (PI) of groups MCF-7/CCL5-siRNA, MCF-7/CCL5-N and MCF-7 were 0.48 ± 0.03, 0.43 ± 0.01 and 0.45 ± 0.02. The PI of groups MDA-MB-231/CCL5-siRNA, MDA-MB-231/CCL5-N and MDA-MB-231 cells were 0.48 ± 0.02, 0.44 ± 0.05 and 0.47 ± 0.02. There was no statistical difference among them (P > 0.05).
  
  
  CONCLUSIONS
  The down-regulation of CCL5 gene in human breast cancer cells may significantly suppress their colony formation ability, rather than affecting their population doubling time to some extent.
authors.affiliation: The Second Department of Gynecology, Renmin Hospital of Wuhan University, Wuhan 430060, China.
authors.forename: Jun-xiu
authors.initials: JX
authors.lastname: Kuang
authors.forename: Wei-xing
authors.initials: WX
authors.lastname: Wang
authors.forename: Sheng-rong
authors.initials: SR
authors.lastname: Sun
authors.forename: Wan-rong
authors.initials: WR
authors.lastname: Wang
authors.forename: Xiao-li
authors.initials: XL
authors.lastname: Yao
mesh_terms: D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018946:Chemokine CCL5 / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D015536:Down-Regulation; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
chemical_list: C483024:CCL5 protein, human; D018946:Chemokine CCL5; D012333:RNA, Messenger; D034741:RNA, Small Interfering