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Logo of bmccancBioMed Centralsearchsubmit a manuscriptregisterthis articleBMC Cancer
 
BMC Cancer. 2012; 12: 370.
Published online Aug 27, 2012. doi:  10.1186/1471-2407-12-370
PMCID: PMC3493279
Knock-down of NDRG2 sensitizes cervical cancer Hela cells to cisplatin through suppressing Bcl-2 expression
Junye Liu,#1,2 Le Yang,#2,3 Jian Zhang,2 Jing Zhang,2 Yongbin Chen,1 Kangchu Li,1 Yurong Li,1 Yan Li,2 Libo Yao,corresponding author2 and Guozhen Guocorresponding author1
1Department of Radiation Medicine, Fourth Military Medical University, Xi’an, China
2Department of Biochemistry and Molecular Biology, State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xi’an, China
3Department of Oncology, Xi’an Central Hospital, Xi’an, China
corresponding authorCorresponding author.
#Contributed equally.
Junye Liu: junyeliu/at/fmmu.edu.cn; Le Yang: leyang79/at/gmail.com; Jian Zhang: biozhangj/at/yahoo.com.cn; Jing Zhang: zhangjing.99/at/163.com; Yongbin Chen: chenyongbin/at/fmmu.edu.cn; Kangchu Li: kangchu/at/fmmu.edu.cn; Yurong Li: 722lyr/at/fmmu.edu.cn; Yan Li: liyanfmmu/at/yahoo.com; Libo Yao: bioyao/at/fmmu.edu.cn; Guozhen Guo: guozhen/at/fmmu.edu.cn
Received February 1, 2012; Accepted August 22, 2012.
Abstract
Background
NDRG2, a member of N-Myc downstream regulated gene family, plays some roles in cellular stress, cell differentiation and tumor suppression. We have found that NDRG2 expression in cervical cancer Hela cells increases significantly upon stimulation with cisplatin, the most popular chemotherapeutic agent currently used for the treatment of advanced cervical cancer. This interesting phenomenon drove us to evaluate the role of NDRG2 in chemosensitivity of Hela cells.
Methods
In the present study, RNA interference was employed to down-regulate NDRG2 expression in Hela cells. RT-PCR and Western blot were used to detect expression of NDRG2, Bcl-2 and Bax in cancer cells. Real-time PCR was applied to detect miR-15b and miR-16 expression levels. Drug sensitivity was determined with MTT assay. Cell cloning efficiency was evaluated by Colony-forming assay. Apoptotic cells were detected with annexin V staining and flow cytometry.
Results
In vitro drug sensitivity assay revealed that suppression of NDRG2 could sensitize Hela cells to cisplatin. Down-regulation of NDRG2 didn’t influence the colony-forming ability but promoted cisplatin-induced apoptosis of Hela cells. Inhibition of NDRG2 in Hela cells was accompanied by decreased Bcl-2 protein level. However, Bcl-2 mRNA level was not changed in Hela cells with down-regulation of NDRG2. Further study indicated that miR-15b and miR-16, two microRNAs targetting Bcl-2, were significantly up-regulated in NDRG2-suppressed Hela cells.
Conclusions
These data suggested that down-regulation of NDRG2 could enhance sensitivity of Hela cells to cisplatin through inhibiting Bcl-2 protein expression, which might be mediated by up-regulating miR-15b and miR-16.
Keywords: Bcl-2, Chemosensitivity, Cisplatin, NDRG2, RNA interference
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