title: [RNA interference silencing expression of survivin gene and reversing drug resistance of ovarian cancer cell line SKOV3/ADM].
abstract: OBJECTIVE
  To examine expression of survivin gene in ovarian epithelial carcinoma drug resistant cell line SKOV3/ADM and its parental cell line SKOV3, and induction of cells apoptosis and reversal of drug resistance in SKOV3/ADM after RNA interference (RNAi) silencing survivin gene.
  
  
  METHODS
  The transcription of survivin gene in cells was detected by semi-quantitative RT-PCR, the protein expression level of survivin gene was analyzed by immunofluorescence staining. SKOV3/ADM cells were treated with pshRNA-survivin and paclitaxel (Taxol), and acridine orange (AO)/ethidium bromide (EB) staining was performed to evaluate the apoptosis of cells.
  
  
  RESULTS
  Survivin gene mRNA expressed by 99.1% and 75.3% respectively in cell lines SKOV3/ADM and SKOV3, while fluorescent cells were 59 +/- 5 and 42 +/- 3 (P < 0.05). After the introduction of pshRNA-survivin into SKOV3/ADM, mRNA transcription level of survivin gene decreased distinctly from 99.1% to 7.9%. The apoptotic cells of control group detected by AO/EB staining was 3.6 +/- 0.6, of Taxol group 10.2 +/- 1.0, of RNAi group 48.5 +/- 4.9, of RNAi + Taxol group 71.5 +/- 6.8. Apoptosis ratio between RNAi + Taxol group and RNAi group had significant difference (P < 0.05), and that between RNAi + Taxol group and Taxol group also had significant difference (P < 0.05).
  
  
  CONCLUSIONS
  Both survivin gene mRNA and its protein are over-expressed in ovarian epithelial carcinoma cell lines SKOV3 and SKOV3/ADM, the level of survivin gene expressed in SKOV3/ADM is obviously different compared with that in its parental cell line SKOV3. RNA interference targeted against specific sequences of survivin in SKOV3/ADM cell could significantly reduce the level of survivin mRNA transcripts and protein, effectively induce the cells apoptosis and restore the sensitivity of cell to conventional chemotherapeutic agents Taxol.
authors.affiliation: Department of Obstetrics and Gynecology, Second Affiliated Hospital, Chongqing University of Medical Science, Chongqing 400010, China.
authors.forename: Kai-xian
authors.initials: KX
authors.lastname: Deng
authors.forename: Ling
authors.initials: L
authors.lastname: Zhong
authors.forename: Mei-xian
authors.initials: MX
authors.lastname: Jiang
authors.forename: Ping-ling
authors.initials: PL
authors.lastname: Wang
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D005260:Female; D020868:Gene Silencing; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis / Q000235:genetics*; D009363:Neoplasm Proteins / Q000235:genetics*; D010051:Ovarian Neoplasms / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077022:Survivin; D014162:Transfection
chemical_list: D000970:Antineoplastic Agents; C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D000077022:Survivin