[Suppression of WWOX gene by RNA interference reverses platinum resistance acquired in SKOV3/SB cells].
OBJECTIVE
To assess the suppression effect on WWOX gene in SKOV3/SB cell line by small interference RNA (siRNA).


METHODS
Transfection of siRNA using lipofectamine 2000 was conducted to silence WWOX gene expression, the expression levels of WWOX mRNA and protein were evaluated, and the effects on the cell cycles at 48 hours of transfection were assessed by RT-PCR, western blot and flow cytometry (FCM) respectively. The cisplatin resistance index was assayed after transfection of SKOV3/SB with siRNA by methyl thiazolyl tetrazolium (MTT) and the cisplatin concentration of SKOV3/SB cells transfected with siRNA of WWOX was measured by high performance liquid chromatography.


RESULTS
(1) In SKOV3/SB cells transfected with WWOX interference fragment, whether at the mRNA or protein level, the expression of both of WWOX decreased. There was a significant difference (P < 0.05) compared with SKOV3 cells and non-transfected cells. (2) After transfection of the WWOX interference fragment, the index of platinum resistance of SKOV3/SB decreased from 5.04 to 3.89. (3) The number of cell transfected with the WWOX interference fragment in G(1) phase was increased, while that in S-phase was decreased. (4) The cisplatin concentration of SKOV3/SB cells transfected with the WWOX interference fragment was increased from 9.43 ng/L to 23.45 ng/L compared with SKOV3/SB cells non-transfected with a significant difference (P < 0.05).


CONCLUSIONS
WWOX gene may be involved in cisplatin resistance phenomenon in epithelial ovarian cancer.
Liu
Yuan-Yuan
YY
Department of Gynecologic Oncology, Tumor Hospital, Guangxi Medical University, Nanning 530021, China.
Li
Li
L
Li
Dan-Rong
DR
Zhang
Wei
W
Wang
Qi
Q
D000970:Antineoplastic Agents / Q000494:pharmacology; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology*; D015536:Down-Regulation; D019008:Drug Resistance, Neoplasm; D005260:Female; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D010051:Ovarian Neoplasms / Q000378:metabolism* / Q000473:pathology; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000235:genetics* / Q000378:metabolism; D000074584:WW Domain-Containing Oxidoreductase
D000970:Antineoplastic Agents; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D025521:Tumor Suppressor Proteins; D010088:Oxidoreductases; D000074584:WW Domain-Containing Oxidoreductase; C486071:WWOX protein, human; D002945:Cisplatin