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  • Small Interfering RNA (siRNA)-Mediated Silencing of the M2 Subunit of Ribonucleotide Reductase: A Novel Therapeutic Strategy in Ovarian Cancer

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Basic Science
Small Interfering RNA (siRNA)-Mediated Silencing of the M2 Subunit of Ribonucleotide Reductase: A Novel Therapeutic Strategy in Ovarian Cancer
  1. Meng Zhang, MMSc*,
  2. Jing Wang, BMSc†,
  3. RuYong Yao, MD‡ and
  4. LiMing Wang, MD, PhD§
  1. *Qingdao University, Qingdao, China;
  2. †Department of Obstetrics and Gynecology, the Hospital of Kenli, Dongying, China;
  3. ‡Department of Central Laboratory, the Affiliated Hospital of Medical College of Qingdao University, Qingdao, China;
  4. §Department of Obstetrics and Gynecology, the Affiliated Hospital of Medical College of Qingdao University, Qingdao, China.
  1. Address correspondence and reprint requests to LiMing Wang, PhD, Department of Obstetrics and Gynecology, the Affiliated Hospital of Medical College of Qingdao University, Qingdao, Shandong 266003, China. E-mail: wlmqingyi@163.com.

Abstract

Objective To investigate the effects of small interfering RNA (siRNA)-mediated silencing of the ribonucleotide reductase M2 subunit (RRM2) on the apoptosis and the drug sensitivity of cisplatin-resistant SKOV3/DDP cells.

Methods Small interfering RNA transfection was mediated by lipofectamine 2000 to silence RRM2 gene. Messenger RNA (mRNA), and protein expression levels of RRM2 were evaluated by real-time polymerase chain reaction and Western blot after transfection. The cell growth inhibition rate was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. The cellular apoptosis and cycling was identified by flow cytometry (FCM).

Results The messenger RNA and protein expression levels of RRM2 markedly decreased after the RRM2 siRNA transfection. The half inhibition concentration of cisplatin in RRM2-RNA interference cells (interference group) was lower than that in RRM2-negative cells (noninterference group) and the SKOV3/DDP cells (blank control group) (P = 0.032). Small interfering RNA–mediated inhibition of RRM2 effectively induced G1/S-phase cell cycle arrest and increased drug (gemcitabine and cisplatin)-induced apoptotic fraction at 72 hours ( (96% ± 3.0)%) after transfection (P < 0.05).

Conclusion Small interfering RNA–mediated RRM2 knockdown significantly reversed SKOV3/DDP cell resistance to cisplatin. RNA interference technology combined with gemcitabine and cisplatin can effectively improve the apoptosis rate of the cisplatin-resistant ovarian cancer cell, which is expected to become the first-line treatment options for the cisplatin-resistant ovarian cancer.

  • Ovarian cancer
  • Gene silencing
  • RNA interference
  • Ribonucleotide reductase M2
  • Cisplatin resistance
  • Apoptosis

https://doi.org/10.1097/IGC.0b013e318287e2b3

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    Footnotes

    • This study was supported by grants from Application and Development Funds of Beijing Municipal Science & Technology Commission (No. 11-2-4-2-(10)-jch).

    • The authors declare no conflicts of interest.

    • Supplemental digital content is available for this article. Direct URL citation appears in the printed text and is provided in the HTML and PDF versions of this article on the journal’s Web site (www.ijgc.net).