MiR-339 and especially miR-766 reactivate the expression of tumor suppressor genes in colorectal cancer cell lines through DNA methyltransferase 3B gene inhibition.
It is observed that upregulation of DNMT3B enzyme in some cancers, including colon cancer, could lead to silencing of tumor suppressor genes. MiR-339 and miR-766 have been predicted to target 3'UTR of DNMT3B gene. Luciferase reporter assay validated that individual and co-transfection of miR-766 and miR-339 into the HEK293T cell reduced luciferase activity to 26% ± 0.41%, 43% ± 0.42 and 64% ± 0.52%, respectively, compared to the control (P < 0.05). Furthermore, transduction of miR-339 and miR-766 expressing viruses into colon cancer cell lines (SW480 and HCT116) decreased DNMT3B expression (1.5, 3-fold) and (3, 4-fold), respectively. In addition, DNA methylation of some tumor suppressor genes decreased. Expression of these genes such as SFRP1 (2 and 1.6-fold), SFRP2 (0.07 and 4-fold), WIF1 (0.05 and 4-fold), and DKK2 (2 and 4-fold) increased in SW-339 and SW-766 cell lines; besides, expression increments for these genes in HCT-339 and HCT-766 cell lines were (2.8, 4-fold), (0.005, 1.5-fold), (1.7 and 3-fold) and (0.04, 1.7-fold), respectively. Also, while in SW-766, cell proliferation reduced to 2.8% and 21.7% after 24 and 48 hours, respectively, SW-339 showed no reduced proliferation. Meanwhile, HCT-766 and HCT-339 showed (3.5%, 12.8%) and (18.8%, 33.9%) reduced proliferation after 24 and 48 hours, respectively. Finally, targeting DNMT3B by these miRs, decreased methylation of tumor suppressor genes such as SFRP1, SFRP2, WIF1 and DKK2 in the mentioned cell lines, and returned the expression of these tumor suppressor genes which can contribute to lethal effect on colon cancer cells and reducing tumorigenicity of these cells.
a Molecular Medicine Department , Pasteur Institute of Iran , Tehran , Iran.
Ali
A
Afgar
b Department of Biochemistry , Pasteur Institute of Iran , Tehran , Iran.
Pezhman
P
Fard-Esfahani
a Molecular Medicine Department , Pasteur Institute of Iran , Tehran , Iran.
Amirhosein
A
Mehrtash
c Virology Department , Pasteur Institute of Iran , Tehran , Iran.
Kayhan
K
Azadmanesh
a Molecular Medicine Department , Pasteur Institute of Iran , Tehran , Iran.
Farnaz
F
Khodarahmi
a Molecular Medicine Department , Pasteur Institute of Iran , Tehran , Iran.
Mahdis
M
Ghadir
a Molecular Medicine Department , Pasteur Institute of Iran , Tehran , Iran.
Ladan
L
Teimoori-Toolabi
D048868:Adaptor Proteins, Signal Transducing / Q000096:biosynthesis / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015179:Colorectal Neoplasms / Q000201:enzymology / Q000235:genetics*; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D016147:Genes, Tumor Suppressor; D045325:HCT116 Cells; D057809:HEK293 Cells; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis / Q000235:genetics; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D000097584:DNA Methyltransferase 3B; D000094742:Secreted Frizzled-Related Proteins
D048868:Adaptor Proteins, Signal Transducing; D004248:DNA (Cytosine-5-)-Methyltransferases; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D035683:MicroRNAs; D012097:Repressor Proteins; D000097584:DNA Methyltransferase 3B; C498622:DKK2 protein, human; C543820:MIRN339 microRNA, human; C575913:MIRN766 microRNA, human; C117636:SFRP1 protein, human; D000094742:Secreted Frizzled-Related Proteins; C109618:SFRP2 protein, human; C485569:WIF1 protein, human
Colorectal neoplasms; DNA methyltransferase 3B; genes; methylation; microRNAs; tumor suppressor
Lab Invest. 2012 Jul;92(7):1033-44
22508389
Cancer Cell. 2006 Mar;9(3):189-98
16530703
Nat Rev Mol Cell Biol. 2010 Apr;11(4):252-63
20216554
Oncol Lett. 2014 Aug;8(2):719-725
25009651
Carcinogenesis. 2008 Mar;29(3):579-84
18192692
Blood. 2009 Jun 18;113(25):6411-8
19211935
Anticancer Drugs. 2015 Feb;26(2):187-96
25325304
Adv Nutr. 2015 Jan 15;6(1):134-5
25593153
Cancer Biother Radiopharm. 2010 Feb;25(1):105-16
20187803
Exp Mol Pathol. 2015 Apr;98(2):200-7
25746661
Bratisl Lek Listy. 2014;115(10):611-6
25573726
Mol Cancer. 2008 Jan 25;7:15
18221536
Gut. 2005 Sep;54(9):1283-6
15843421
Carcinogenesis. 2007 Dec;28(12):2434-42
17893234
Nucleic Acids Res. 1999 Jun 1;27(11):2291-8
10325416
Am J Pathol. 2007 Sep;171(3):728-38
17724137
Hum Mol Genet. 2001 Jun 15;10(13):1413-9
11440994
Clin Infect Dis. 2015 Feb 15;60(4):627-38
25362204
Nat Genet. 2005 May;37(5):495-500
15806104
Proc Natl Acad Sci U S A. 2007 Oct 2;104(40):15805-10
17890317
Genome Res. 2009 Jan;19(1):92-105
18955434
J Clin Invest. 2012 Jan;122(1):163-77
22133874
Stroke. 2015 May;46(5):e121-2
25873596
Clin Cancer Res. 2009 Jun 1;15(11):3663-71
19470733
Cancer Biol Ther. 2005 May;4(5):585-92
15908783
Oncogene. 2005 Nov 24;24(53):7946-52
16007117
Gut. 2011 Apr;60(4):499-508
21068132
RNA. 2008 May;14(5):872-7
18367714
Circulation. 2013 May 28;127(21):2075-7
23625958
J Neurooncol. 2011 Sep;104(2):483-94
21229291
Cancer Biol Ther. 2014 Apr;15(4):419-27
24448385
Nature. 2008 Mar 6;452(7183):45-50
18322525
Mod Pathol. 2007 Jul;20(7):711-21
17464311
Int J Oncol. 2012 Aug;41(2):721-32
22664488
Oncogenesis. 2013 Sep 16;2:e71
24042735
Nat Rev Gastroenterol Hepatol. 2011 Oct 18;8(12):686-700
22009203
Mol Carcinog. 2007 Sep;46(9):766-72
17538945
Cell Cycle. 2008 Oct;7(20):3143-8
18843198
Genes Dev. 2007 Dec 1;21(23):3110-22
18056424
J Zhejiang Univ Sci B. 2008 Jan;9(1):1-4
18196605
Br J Cancer. 2006 Nov 20;95(10):1415-8
17003783
Am J Pathol. 2003 Apr;162(4):1361-71
12651628
Adv Genet. 2010;70:87-99
20920746
Mol Cell Biol. 2002 May;22(9):3157-73
11940673
BMC Cancer. 2014 Nov 29;14:891
25432628