Top

Journal of Experimental & Clinical Cancer Research

Published in:

Open Access 01-12-2011 | Research

Effects of RNA interference-mediated gene silencing of JMJD2A on human breast cancer cell line MDA-MB-231 in vitro

Authors: Bei-Xu Li, Ming-Chang Zhang, Cheng-Liang Luo, Peng Yang, Hui Li, Hong-Mei Xu, Hong-Fei Xu, Yi-Wen Shen, Ai-Min Xue, Zi-Qin Zhao

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2011

Abstract

Previous data demonstrate that JMJD2A is a cancer-associated gene and may be involved in human breast cancer by demethylation of H3K9me3. The aim of this study was to investigate depressive effects on JMJD2A by transfection with JMJD2A-sepcific siRNA in human breast cancer cell line MDA-MB-231 and effects on cell proliferation, invasion and migration. JMJD2A-specific siRNA was chemically synthesised and transfected into human breast cancer cell line MDA-MB-231. Expression levels of JMJD2A were detected by quantitative real-time PCR and Western blot analysis. Cells proliferation was evaluated by using flow cytometric anlysis and MTT assay. The abilities of invasion and migration were evaluated by cell migration and invasion assay with Boyden chambers. The results showed that the transfection was successful and expression levels of JMJD2A mRNA and protein in siRNA group were both down-regulated. By MTT assay, the mean actual absorbance in siRNA group was significantly lower than that in blank control group (P < 0.05) and negative control group (P < 0.05). In addition, the percentage of cells in G0/G1 phase in siRNA group was significantly more than that in blank control group (P < 0.05) and negative control group (P < 0.05). Furthermore, by cell invasion and migration assay, the decreased number of migrated cells in siRNA group was observed (P < 0.05). These data imply that silencing JMJD2A gene could result in cell cycle change and proliferation inhibition, and lead to suppress tumor cell invasion and migration. It provides a new perspective in understanding the pleiotropic functions of JMJD2A and its contribution to human breast cancer.

Available only for authorised users

Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D: Global cancer statistics. CA Cancer J Clin CA Cancer J Clin. 2011, 61: 69-90.CrossRefPubMed

Sen GL, Blau HM: A brief history of RNAi: the silence of the genes. FASEB J. 2006, 20: 1293-1299. 10.1096/fj.06-6014rev.CrossRefPubMed

Katoh M, Katoh M: Identification and characterization of JMJD2 family genes in silico. Int J Oncol. 2004, 24: 1623-1628.PubMed

Trojer P, Reinberg D: Histone lysine demethylases and their impact on epigenetics. Cell. 2006, 125: 213-217. 10.1016/j.cell.2006.04.003.CrossRefPubMed

Whetstine JR, Nottke A, Lan F, Huarte M, Smolikov S, Chen Z, Spooner E, Li E, Zhang G, Colaiacovo M, Shi Y: Reversal of Histone Lysine Trimethylation by the JMJD2 Family of Histone Demethylases. Cell. 2006, 125: 467-481. 10.1016/j.cell.2006.03.028.CrossRefPubMed

Nottke A, Colaiácovo MP, Shi Y: Developmental roles of the histone lysine demethylases. Development. 2009, 136: 879-889. 10.1242/dev.020966.PubMedCentralCrossRefPubMed

Gray SG, Iglesias AH, Lizcano F, Villanueva R, Camelo S, Jingu H, Teh BT, Koibuchi N, Chin WW, Kokkotou E, Dangond F: Functional Characterization of JMJD2A, a Histone Deacetylase- and Retinoblastoma-binding Protein. J Biol Chem. 2005, 280: 28507-28518. 10.1074/jbc.M413687200.CrossRefPubMed

Shin S, Janknecht R: Activation of androgen receptor by histone demethylases JMJD2A and JMJD2D. Biochem Biophys Res Commun. 2007, 359: 742-746. 10.1016/j.bbrc.2007.05.179.CrossRefPubMed

Zhang XD, Wang Y, Wang Y, Zhang X, Han R, Wu JC, Liang ZQ, Gu ZL, Han F, Fukunaga K, Qin ZH: p53 mediates mitochondria dysfunction-triggered autophagy activation and cell death in rat striatum. Autophagy. 2009, 5: 339-350. 10.4161/auto.5.3.8174.CrossRefPubMed

10.

Luo CL, Li BX, Li QQ, Chen XP, Sun YX, Bao HJ, Dai DK, Shen YW, Xu HF, Ni H, Wan L, Qin ZH, Tao LY, Zhao ZQ: Autophagy is involved in traumatic brain injury-induced cell death and contributes to functional outcome deficits in mice. Neuroscience. 2011, 184: 54-63.CrossRefPubMed

11.

Dai HY, Liu L, Qin SK, He XM, Li SY: Lobaplatin suppresses proliferation and induces apoptosis in the human colorectal carcinoma cell Line LOVO in vitro. Biomed Pharmacother. 2011, 65: 137-141. 10.1016/j.biopha.2010.12.001.CrossRefPubMed

12.

Li L, Zhang C, Li X, Lu S, Zhou Y: The candidate tumor suppressor gene ECRG4 inhibits cancer cells migration and invasion in esophageal carcinoma. J Exp Clin Cancer Res. 2010, 29: 133-10.1186/1756-9966-29-133.PubMedCentralCrossRefPubMed

13.

Jovanovic J, Rønneberg JA, Tost J, Kristensen V: The epigenetics of breast cancer. Mol Oncol. 2010, 4: 242-254. 10.1016/j.molonc.2010.04.002.CrossRefPubMed

14.

Martin C, Zhang Y: The diverse functions of histone lysine methylation. Nat Rev Mol Cell Biol. 2005, 6: 838-849. 10.1038/nrm1761.CrossRefPubMed

15.

Müller-Tidow C, Klein HU, Hascher A, Isken F, Tickenbrock L, Thoennissen N, Agrawal-Singh S, Tschanter P, Disselhoff C, Wang Y, Becker A, Thiede C, Ehninger G, zur Stadt U, Koschmieder S, Seidl M, Müller FU, Schmitz W, Schlenke P, McClelland M, Berdel WE, Dugas M, Serve H, Study Alliance Leukemia: Profiling of histone H3 lysine 9 trimethylation levels predicts transcription factor activity and survival in acute myeloid leukemia. Blood. 2010, 116: 3564-3571. 10.1182/blood-2009-09-240978.PubMedCentralCrossRefPubMed

16.

Cloos PA, Christensen J, Agger K, Helin K: Erasing the methyl mark: histone demethylases at the center of cellular differentiation and disease. Genes Dev. 2008, 22: 1115-1140. 10.1101/gad.1652908.PubMedCentralCrossRefPubMed

17.

Peters AH, O'Carroll D, Scherthan H, Mechtler K, Sauer S, Schöfer C, Weipoltshammer K, Pagani M, Lachner M, Kohlmaier A, Opravil S, Doyle M, Sibilia M, Jenuwein T: Loss of the Suv39h histone methyltransferases impairs mammalian heterochromatin and genome stability. Cell. 2001, 107: 323-337. 10.1016/S0092-8674(01)00542-6.CrossRefPubMed

18.

Braig M, Lee S, Loddenkemper C, Rudolph C, Peters AH, Schlegelberger B, Stein H, Dörken B, Jenuwein T, Schmitt CA: Oncogene-induced senescence as an initial barrier in lymphoma development. Nature. 2005, 436: 660-665. 10.1038/nature03841.CrossRefPubMed

19.

Schübeler D, MacAlpine DM, Scalzo D, Wirbelauer C, Kooperberg C, van Leeuwen F, Gottschling DE, O'Neill LP, Turner BM, Delrow J, Bell SP, Groudine M: The histone modification pattern of active genes revealed through genome-wide chromatin analysis of higher eukaryote. Genes Dev. 2004, 18: 1263-1271. 10.1101/gad.1198204.PubMedCentralCrossRefPubMed

20.

Shilatifard A: Chromatin modifications by methylation and ubiquitination: implications in the regulation of gene expression. Annu Rev Biochem. 2006, 75: 243-269. 10.1146/annurev.biochem.75.103004.142422.CrossRefPubMed

21.

Xu D, Bai J, Duan Q, Costa M, Dai W: Covalent modifications of histones during mitosis and meiosis. Cell Cycle. 2009, 8: 3688-3694. 10.4161/cc.8.22.9908.CrossRefPubMed

22.

Mikkelsen TS, Ku M, Jaffe DB, Issac B, Lieberman E, Giannoukos G, Alvarez P, Brockman W, Kim TK, Koche RP, Lee W, Mendenhall E, O'Donovan A, Presser A, Russ C, Xie X, Meissner A, Wernig M, Jaenisch R, Nusbaum C, Lander ES, Bernstein BE: Genome-wide maps of chromatin state in pluripotent and lineage-committed cells. Nature. 2007, 448: 553-560. 10.1038/nature06008.PubMedCentralCrossRefPubMed

23.

Barski A, Cuddapah S, Cui K, Roh TY, Schones DE, Wang Z, Wei G, Chepelev I, Zhao K: High-resolution profiling of histone methylations in the human genome. Cell. 2007, 129: 823-837. 10.1016/j.cell.2007.05.009.CrossRefPubMed

24.

Brinkman AB, Roelofsen T, Pennings SW, Martens JH, Jenuwein T, Stunnenberg HG: Histone modification patterns associated with the human X chromosome. EMBO Rep. 2006, 7: 628-634.PubMedCentralPubMed

25.

Vakoc CR, Mandat SA, Olenchock BA, Blobel GA: Histone H3 lysine 9 methylation and HP1gamma are associated with transcription elongation through mammalian chromatin. Mol Cell. 2005, 19: 381-391. 10.1016/j.molcel.2005.06.011.CrossRefPubMed

26.

Gomes NP, Espinosa JM: Gene-specific repression of the p53 target gene PUMA via intragenic CTCF-Cohesin binding. Genes Dev. 2010, 24: 1022-1034. 10.1101/gad.1881010.PubMedCentralCrossRefPubMed

27.

Kawazu M, Saso K, Tong KI, McQuire T, Goto K, Son DO, Wakeham A, Miyagishi M, Mak TW, Okada H: Histone demethylase JMJD2B functions as a co-factor of estrogen receptor in breast cancer proliferation and mammary gland development. PLoS One. 2011, 6: e17830-10.1371/journal.pone.0017830.PubMedCentralCrossRefPubMed

28.

Gray SG, Iglesias AH, Lizcano F, Villanueva R, Camelo S, Jingu H, Teh BT, Koibuchi N, Chin WW, Kokkotou E, Dangond F: Functional characterization of JMJD2A, a histone deacetylase- and retinoblastoma-binding protein. J Biol Chem. 2005, 280: 28507-28518. 10.1074/jbc.M413687200.CrossRefPubMed

29.

Takaki T, Fukasawa K, Suzuki-Takahashi I, Hirai H: Cdk-mediated phosphorylation of pRB regulates HDAC binding in vitro. Biochem Biophys Res Commun. 2004, 316: 252-255. 10.1016/j.bbrc.2004.02.044.CrossRefPubMed

30.

Lai A, Kennedy BK, Barbie DA, Bertos NR, Yang XJ, Theberge MC, Tsai SC, Seto E, Zhang Y, Kuzmichev A, Lane WS, Reinberg D, Harlow E, Branton PE: RBP1 recruits the mSIN3-histone deacetylase complex to the pocket of retinoblastoma tumor suppressor family proteins found in limited discrete regions of the nucleus at growth arrest. Mol Cell Biol. 2001, 21: 2918-2932. 10.1128/MCB.21.8.2918-2932.2001.PubMedCentralCrossRefPubMed

31.

Yu Y, Xu F, Peng H, Fang X, Zhao S, Li Y, Cuevas B, Kuo WL, Gray JW, Siciliano M, Mills GB, Bast RC: NOEY2 (ARHI), an imprinted putative tumor suppressor gene in ovarian and breast carcinomas. Proc Natl Acad Sci USA. 1999, 96: 214-219. 10.1073/pnas.96.1.214.PubMedCentralCrossRefPubMed

32.

Lu Z, Luo RZ, Peng H, Huang M, Nishmoto A, Hunt KK, Helin K, Liao WS, Yu Y: E2F-HDAC complexes negatively regulate the tumor suppressor gene ARHI in breast cancer. Oncogene. 2006, 25: 230-239.CrossRefPubMed

Title: Effects of RNA interference-mediated gene silencing of JMJD2A on human breast cancer cell line MDA-MB-231 in vitro
Authors: Bei-Xu Li
Ming-Chang Zhang
Cheng-Liang Luo
Peng Yang
Hui Li
Hong-Mei Xu
Hong-Fei Xu
Yi-Wen Shen
Ai-Min Xue
Zi-Qin Zhao
Publication date: 01-12-2011
Publisher: BioMed Central
Published in: Journal of Experimental & Clinical Cancer Research / Issue 1/2011
Electronic ISSN: 1756-9966
DOI: https://doi.org/10.1186/1756-9966-30-90

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

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 1/2011

Polymorphisms in the SULF1 gene are associated with early age of onset and survival of ovarian cancer

Expression of Ets-1, Ang-2 and maspin in ovarian cancer and their role in tumor angiogenesis

Factors that contribute to long-term survival in patients with leukemia not in remission at allogeneic hematopoietic cell transplantation

Adjuvant chemotherapy for resected non-small-cell lung cancer: future perspectives for clinical research

Effects of DNMT1 silencing on malignant phenotype and methylated gene expression in cervical cancer cells

The association of Streptococcus bovis/gallolyticus with colorectal tumors: The nature and the underlying mechanisms of its etiological role

Keynote webinar | Spotlight on antibody–drug conjugates in cancer