title: [Role of microRNA-21 in the proliferation and apoptosis of ovarian epithelial carcinoma cells].
abstract: OBJECTIVE
  To investigate the role and mechanism of microRNA-21(miR-21) in the proliferation and apoptosis of ovarian epithelial carcinoma cells.
  
  
  METHODS
  A short-hairpin RNA specifically targeting miR-21 plasmid was constructed, and the recombinant was identified by restriction endonuclease analysis and DNA sequencing. Three experimental groups were included, transfection group (transfected with pSIREN-miR-21), negative control group (transfected with pSIREN-miR-21-neg) and blank control group (without transfection plasmid). The expression of miR-21 was detected by stem-loop real-time reverse transcription (RT)-PCR in OVCAR3 cells, and western blot was used to detect the expression of programmed cell death 4 (PDCD4) protein. Tethyl thiazolyl tetrazolium(MTT) and flow cytometry method were used respectively.
  
  
  RESULTS
  Recombinant plasmid (pSIREN-miR-21) was constructed successfully and identified by restriction endonuclease analysis and DNA sequencing. The relative expression level of miR-21 in cells transfection, negative control and blank control group was 0.26 ± 0.08, 1.26 ± 0.21 and 1.00 respectively. The level of miR-21 in the cells in transfection group was significantly lower than those in the negative control and blank control group (P < 0.01). The gray scale of PDCD4 protein was 1443 ± 33, 858 ± 19 and 846 ± 16 in the transfection group, negative control and blank control group respectively. The value of PDCD4 in transfection group was higher than other control groups, and there were significantly difference among them(P < 0.01). Moreover, the optical density of the cells in transfection group was 0.661 ± 0.015, significantly lower than those in two control groups (0.848 ± 0.150 for negative control, 0.935 ± 0.133 for blank control, P < 0.01). Forty-eight hours after transfection, the rate of viable apoptotic cell was significantly higher than negative control and blank control group [(25.821 ± 0.763)% vs. (0.010 ± 0.003)% vs. (0.238 ± 0.023)%; P < 0.01]; 72 hours after transfection, the rates of viable apoptotic cell and necrotic cell were all higher than the two control groups [the rate of viable apoptotic cell was (30.480 ± 0.821)%, (7.792 ± 0.312)% and (7.033 ± 0.257)% respectively (P < 0.01); the rate of necrotic cell was (3.558 ± 0.211)%, (1.557 ± 0.067)% and (1.049 ± 0.028)%, respectively (P < 0.01)].
  
  
  CONCLUSIONS
  miR-21 might play an important role in the proliferation and apoptosis of ovarian epithelial carcinoma cells through negatively control the expression of PDCD4.
authors.affiliation: Department of Gynecology, Qingdao University, Qingdao, China. lyh7497@163.com
authors.forename: Yan-hui
authors.initials: YH
authors.lastname: Lou
authors.forename: Zhu-mei
authors.initials: ZM
authors.lastname: Cui
authors.forename: Fu-ling
authors.initials: FL
authors.lastname: Wang
authors.forename: Xing-sheng
authors.initials: XS
authors.lastname: Yang
mesh_terms: D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D000077216:Carcinoma, Ovarian Epithelial; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015536:Down-Regulation / Q000235:genetics; D005260:Female; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D009375:Neoplasms, Glandular and Epithelial / Q000235:genetics / Q000378:metabolism*; D010051:Ovarian Neoplasms / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D034741:RNA, Small Interfering / Q000235:genetics*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D011995:Recombination, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
chemical_list: D051017:Apoptosis Regulatory Proteins; C524940:MIRN21 microRNA, human; D035683:MicroRNAs; C114487:PDCD4 protein, human; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins