Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress 2023 EHA Congress 2023 ASCO Annual Meeting
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
BMC Cancer
Published in: BMC Cancer 1/2012

Open Access 01-12-2012 | Research article

Knock-down of NDRG2 sensitizes cervical cancer Hela cells to cisplatin through suppressing Bcl-2 expression

Authors: Junye Liu, Le Yang, Jian Zhang, Jing Zhang, Yongbin Chen, Kangchu Li, Yurong Li, Yan Li, Libo Yao, Guozhen Guo

Published in: BMC Cancer | Issue 1/2012

Login to get access

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.
Appendix
Available only for authorised users
Literature
1.
Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM: Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 2010, 127: 2893-2917. 10.1002/ijc.25516.CrossRefPubMed
2.
Green JA, Lainakis G: Cytotoxic chemotherapy for advanced or recurrent cervical cancer. Ann Oncol. 2006, 17: x230-x232. 10.1093/annonc/mdl265.CrossRefPubMed
3.
Jordan P, Carmo-Fonseca M: Molecular mechanisms involved in cisplatin cytotoxicity. Cell Mol Life Sci. 2000, 57: 1229-1235. 10.1007/PL00000762.CrossRefPubMed
4.
Pectasides D, Kamposioras K, Papaxoinis G, Pectasides E: Chemotherapy for recurrent cervical cancer. Cancer Treat Rev. 2008, 34: 603-613. 10.1016/j.ctrv.2008.05.006.CrossRefPubMed
5.
Ozsaran Z, Kamer S, Yalman D, Akagunduz O, Aras A: Treatment results and prognostic factors for cervical cancer patients treated by radiochemotherapy with weekly cisplatin. Eur J Gynaecol Oncol. 2007, 28: 196-200.PubMed
6.
Yao L, Zhang J, Liu X: NDRG2: a Myc-repressed gene involved in cancer and cell stress. Acta Biochim Biophys Sin (Shanghai). 2008, 40: 625-635. 10.1111/j.1745-7270.2008.00434.x.CrossRef
7.
Hu XL, Liu XP, Lin SX, Deng YC, Liu N, Li X, Yao LB: NDRG2 expression and mutation in human liver and pancreatic cancers. World J Gastroenterol. 2004, 10: 3518-3521.CrossRefPubMedPubMedCentral
8.
Lusis EA, Watson MA, Chicoine MR, Lyman M, Roerig P, Reifenberger G, Gutmann DH, Perry A: Integrative genomic analysis identifies NDRG2 as a candidate tumor suppressor gene frequently inactivated in clinically aggressive meningioma. Cancer Res. 2005, 65: 7121-7126. 10.1158/0008-5472.CAN-05-0043.CrossRefPubMed
9.
Choi SC, Yoon SR, Park YP, Song EY, Kim JW, Kim WH, Yang Y, Lim JS, Lee HG: Expression of NDRG2 is related to tumor progression and survival of gastric cancer patients through Fas-mediated cell death. Exp Mol Med. 2007, 39: 705-714.CrossRefPubMed
10.
Lorentzen A, Vogel LK, Lewinsky RH, Saebø M, Skjelbred CF, Godiksen S, Hoff G, Tveit KM, Lothe IM, Ikdahl T, Kure EH, Mitchelmore C: Expression of NDRG2 is down-regulated in high-risk adenomas and colorectal carcinoma. BMC Cancer. 2007, 7: 192-10.1186/1471-2407-7-192.CrossRefPubMedPubMedCentral
11.
Ma J, Jin H, Wang H, Yuan J, Bao T, Jiang X, Zhang W, Zhao H, Yao L: Expression of NDRG2 in clear cell renal cell carcinoma. Biol Pharm Bull. 2008, 31: 1316-1320. 10.1248/bpb.31.1316.CrossRefPubMed
12.
Tepel M, Roerig P, Wolter M, Gutmann DH, Perry A, Reifenberger G, Riemenschneider MJ: Frequent promoter hypermethylation and transcriptional downregulation of the NDRG2 gene at 14q11.2 in primary glioblastoma. Int J Cancer. 2008, 123: 2080-2086. 10.1002/ijc.23705.CrossRefPubMed
13.
Zhao H, Zhang J, Lu J, He X, Chen C, Li X, Gong L, Bao G, Fu Q, Chen S, Lin W, Shi H, Ma J, Liu X, Ma Q, Yao L: Reduced expression of N-Myc downstream-regulated gene 2 in human thyroid cancer. BMC Cancer. 2008, 8: 303-10.1186/1471-2407-8-303.CrossRefPubMedPubMedCentral
14.
Kim YJ, Yoon SY, Kim JT, Song EY, Lee HG, Son HJ, Kim SY, Cho D, Choi I, Kim JH, Kim JW: NDRG2 expression decreases with tumor stages and regulates TCF/beta-catenin signaling in human colon carcinoma. Carcinogenesis. 2009, 30: 598-605. 10.1093/carcin/bgp047.CrossRefPubMedPubMedCentral
15.
Piepoli A, Cotugno R, Merla G, Gentile A, Augello B, Quitadamo M, Merla A, Panza A, Carella M, Maglietta R, D'Addabbo A, Ancona N, Fusilli S, Perri F, Andriulli A: Promoter methylation correlates with reduced NDRG2 expression in advanced colon tumour. BMC Med Genomics. 2009, 2: 11-10.1186/1755-8794-2-11.CrossRefPubMedPubMedCentral
16.
Shi H, Jin H, Chu D, Wang W, Zhang J, Chen C, Xu C, Fan D, Yao L: Suppression of N-myc downstream-regulated gene 2 is associated with induction of Myc in colorectal cancer and correlates closely with differentiation. Biol Pharm Bull. 2009, 32: 968-975. 10.1248/bpb.32.968.CrossRefPubMed
17.
Lorentzen A, Lewinsky RH, Bornholdt J, Vogel LK, Mitchelmore C: Expression profile of the N-myc Downstream Regulated Gene 2 (NDRG2) in human cancers with focus on breast cancer. BMC Cancer. 2011, 11: 14-10.1186/1471-2407-11-14.CrossRefPubMedPubMedCentral
18.
Liu J, Zhang J, Wang X, Li Y, Chen Y, Li K, Zhang J, Yao L, Guo G: HIF-1 and NDRG2 contribute to hypoxia-induced radioresistance of cervical cancer Hela cells. Exp Cell Res. 2010, 316: 1985-1993. 10.1016/j.yexcr.2010.02.028.CrossRefPubMed
19.
Liu N, Wang L, Li X, Yang Q, Liu X, Zhang J, Zhang J, Wu Y, Ji S, Zhang Y, Yang A, Han H, Yao L: N-Myc downstream-regulated gene 2 is involved in p53-mediated apoptosis. Nucleic Acids Res. 2008, 36: 5335-5349. 10.1093/nar/gkn504.CrossRefPubMedPubMedCentral
20.
Wang L, Liu N, Yao L, Li F, Zhang J, Deng Y, Liu J, Ji S, Yang A, Han H, Zhang Y, Zhang J, Han W, Liu X: NDRG2 is a new HIF-1 target gene necessary for hypoxia-induced apoptosis in A549 cells. Cell Physiol Biochem. 2008, 21: 239-250. 10.1159/000113765.CrossRefPubMed
21.
Xia L, Zhang D, Du R, Pan Y, Zhao L, Sun S, Hong L, Liu J, Fan D: miR-15b and miR-16 modulate multidrug resistance by targeting BCL2 in human gastric cancer cells. Int J Cancer. 2008, 123: 372-379. 10.1002/ijc.23501.CrossRefPubMed
22.
Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) Method. Methods. 2001, 25: 402-408. 10.1006/meth.2001.1262.CrossRefPubMed
23.
Zhang HH, Wang XJ, Li GX, Yang E, Yang NM: Detection of let-7a microRNA by real-time PCR in gastric carcinoma. World J Gastroenterol. 2007, 13: 2883-2888.PubMedPubMedCentral
24.
Zhu W, Xu H, Zhu D, Zhi H, Wang T, Wang J, Jiang B, Shu Y, Liu P: miR-200bc/429 cluster modulates multidrug resistance of human cancer cell lines by targeting BCL2 and XIAP. Cancer Chemother Pharmacol. 2012, 69: 723-731. 10.1007/s00280-011-1752-3.CrossRefPubMed
25.
Galluzzi L, Senovilla L, Vitale I, Michels J, Martins I, Kepp O, Castedo M, Kroemer G: Molecular mechanisms of cisplatin resistance. Oncogene. 2012, 31: 1869-1883. 10.1038/onc.2011.384.CrossRefPubMed
26.
Jamieson ER, Lippard SJ: Structure, recognition, and processing of cisplatin-DNA adducts. Chem Rev. 1999, 99: 2467-2498. 10.1021/cr980421n.CrossRefPubMed
27.
Cohen SM, Lippard SJ: Cisplatin: from DNA damage to cancer chemotherapy. Prog Nucleic Acid Res Mol Biol. 2001, 67: 93-130.CrossRefPubMed
28.
Zhang J, Li F, Liu X, Shen L, Liu J, Su J, Zhang W, Deng Y, Wang L, Liu N, Han W, Zhang J, Ji S, Yang A, Han H, Yao L: The repression of human differentiation-related gene NDRG2 expression by Myc via Miz-1-dependent interaction with the NDRG2 core promoter. J Biol Chem. 2006, 281: 39159-39168. 10.1074/jbc.M605820200.CrossRefPubMed
29.
Liu L, Ning X, Sun L, Shi Y, Han S, Guo C, Chen Y, Sun S, Yin F, Wu K, Fan D: Involvement of MGr1-Ag/37LRP in the vincristine-induced HIF-1 expression in gastric cancer cells. Mol Cell Biochem. 2007, 303: 151-160. 10.1007/s11010-007-9467-9.CrossRefPubMed
30.
Hwang J, Kim Y, Kang HB, Jaroszewski L, Deacon AM, Lee H, Choi WC, Kim KJ, Kim CH, Kang BS, Lee JO, Oh TK, Kim JW, Wilson IA, Kim MH: Crystal structure of the human N-Myc downstream-regulated gene 2 protein provides insight into its role as a tumor suppressor. J Biol Chem. 2011, 286: 12450-12460. 10.1074/jbc.M110.170803.CrossRefPubMedPubMedCentral
31.
Liang ZL, Kang K, Yoon S, Huang SM, Lim JS, Kim JM, Lim JS, Lee HJ: NDRG2 Is Involved in the Oncogenic Properties of Renal Cell Carcinoma and Its Loss Is a Novel Independent Poor Prognostic Factor After Nephrectomy. Ann Surg Oncol. 2012, Jan 14. [Epub ahead of print]
Metadata
Title
Knock-down of NDRG2 sensitizes cervical cancer Hela cells to cisplatin through suppressing Bcl-2 expression
Authors
Junye Liu
Le Yang
Jian Zhang
Jing Zhang
Yongbin Chen
Kangchu Li
Yurong Li
Yan Li
Libo Yao
Guozhen Guo
Publication date
01-12-2012
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2012
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-12-370

Other articles of this Issue 1/2012

BMC Cancer 1/2012 Go to the issue

Research article

Time trends of cancer mortality among elderly in Italy, 1970–2008: an observational study

Study protocol

Impact of treatment with bevacizumab beyond disease progression: a randomized phase II study of docetaxel with or without bevacizumab after platinum-based chemotherapy plus bevacizumab in patients with advanced nonsquamous non–small cell lung cancer (WJOG 5910L)

Research article

Identification of PADI2 as a potential breast cancer biomarker and therapeutic target

Research article

Engineered measles virus Edmonston strain used as a novel oncolytic viral system against human hepatoblastoma

Research article

Changed adipocytokine concentrations in colorectal tumor patients and morbidly obese patients compared to healthy controls

Study protocol

Resection of the primary tumour versus no resection prior to systemic therapy in patients with colon cancer and synchronous unresectable metastases (UICC stage IV): SYNCHRONOUS - a randomised controlled multicentre trial (ISRCTN30964555)
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits