Modulation of the proliferation, migration and SIRT1 expression of prostate cancer cell PC-3 by miRNA-221 and miRNA-222

doi:10.3969/j.issn.1673-8640.2014.05.005

Abstract

Abstract:

Objective To investigate the roles of micro RNA(miRNA)-221 and miRNA-222 on the proliferation and migration of prostate cancer cell PC-3 and the influence on silent information regulator 1(SIRT1). Methods The groups were arranged as cells without treatment (control group), cells transfected with pEX-5 empty plasmid (pEX-5 group), cells transfected with miRNA-221 inhibitor (miRNA-221 inhibition group) and cells transfected with miRNA-222 inhibitor (miRNA-222 inhibition group). The influence of miRNA-221 and miRNA-222 on prostate cancer cell proliferation was determined by CCK-8, and the migration ability was determined by wound healing test. The protein and mRNA levels of SIRT1 were determined by Western blotting and fluorescence quantitation polymerase chain reaction (PCR), respectively. The prediction analysis was performed by miRNAanda target. Results After the expression inhibition of miRNA-221 or miRNA-222 in cell PC-3, the cell activities were determined for 7 d. The activities (A_{450 mm}) in miRNA-221 and miRNA-222 inhibition groups were down-regulated about 1.5-2.0 times compared with that in pEX-5 group, and there were significant differences between the 2 groups from the 3rd d(t=6.7, P<0.01; t=5.3, P<0.01). The wound healing test showed that the migration abilities of miRNA-221 inhibition group and miRNA-222 inhibition group decreased than that of pEX-5 group. Moreover, the expression levels of SIRT1 protein in miRNA-221 inhibition group(0.26±0.021) and miRNA-222 inhibition group(0.21±0.005 8) were up-regulated compared with that in pEX-5 group(0.14±0.017), and there were statistical significances (t=6.3 and 6.4, P<0.05 ). The SIRT1 mRNA expression levels had no statistical significance among the 3 groups(P>0.05). Conclusions The miRNA-221 and miRNA-222 promote the proliferation and migration of prostate cancer cell PC-3 and have the influence on SIRT1 expression in protein level.

Key words: Micro RNA-221, Micro RNA-222, Silent information regulator 1, Prostate cancer

CLC Number:

YANG Xiao, GAN Rong, YANG Yingmei, ZHAO Lingxu, BO Jinshuang, GUAN Yanming, MENG Qinghe, L Jianxin.. Modulation of the proliferation, migration and SIRT1 expression of prostate cancer cell PC-3 by miRNA-221 and miRNA-222[J]. , 2014, 29(5): 446-451.

References

[1] Jemal A, Siegel R, Xu J, et al. Cancer statistics, 2010[J]. CA Cancer J Clin, 2011, 61(2): 133-134.
[2] Mitchell PS, Parkin RK, Kroh EM, et al. Circulating microRNAs as stable blood-based markers for cancer detection[J]. Proc Natl Acad Sci USA, 2008, 105(30): 10513-10518.
[3] Feldman BJ, Feldman D. The development of androgen-independent prostate cancer[J]. Nat Rev Cancer, 2001, 1(1): 34-45.
[4] Krol J, Loedige I, Filipowicz W. The widespread regulation of microRNA biogenesis, function and decay[J]. Nat Rev Genet, 2010, 11(9): 597-610.
[5] Belletti B, Nicoloso MS, Schiappacassi M, et al. p27(kip1) functional regulation in human cancer: a potential target for therapeutic designs[J]. Curr Med Chem, 2005, 12(14): 1589-1605.
[6] Chen Y, Zaman MS, Deng G, et al. MicroRNAs 221/222 and genistein-mediated regulation of ARHI tumor suppressor gene in prostate cancer[J]. Cancer Prev Res (Phila), 2011, 4(1): 76-86.
[7] De Boer VC, de Goffau MC, Arts IC, et al. SIRT1 stimulation by polyphenols is affected by their stability and metabolism[J]. Mech Ageing Dev, 2006, 127(7): 618-627.
[8] Wang RH, Sengupta K, Li C, et al. Impaired DNA damage response, genome instability, and tumorigenesis in SIRT1 mutant mice[J]. Cancer Cell, 2008, 14(4): 312-323.
[9] Ciafrè SA, Galardi S, Mangiola A, et al. Extensive modulation of a set of microRNAs in primary glioblastoma[J]. Biochem Biophys Res Commun, 2005, 334(4): 1351-1358.
[10] He H, Jazdzewski K, Li W, et al. The role of microRNA genes in papillary thyroid carcinoma[J]. Proc Natl Acad Sci USA, 2005, 102(52): 19075-19080.
[11] Wu M, Jolicoeur N, Li Z, et al. Genetic variations of microRNAs in human cancer and their effects on the expression of miRNAs[J]. Carcinogenesis, 2008, 29(9): 1710-1716.
[12] Ventura A, Jacks T. MicroRNAs and cancer: short RNAs go a long way[J]. Cell, 2009, 136(4): 586-591.
[13] Galardi S, Mercatelli N, Giorda E, et al. miR-221 and miR-222 expression affects the proliferation potential of human prostate carcinoma cell lines by targeting p27Kip1[J]. J Biol Chem, 2007, 282(32): 23716-23724.
[14] Schaefer A, Jung M, Mollenkopf HJ, et al. Diagnostic and prognostic implications of microRNA profiling in prostate carcinoma[J]. Int J Cancer, 2010, 126(5): 1166-1176.
[15] Miller TE, Ghoshal K, Ramaswamy B, et al. MicroRNA-221/222 confers tamoxifen resistance in breast cancer by targeting p27Kip1[J]. J Biol Chem, 2008, 283(44): 29897-29903.
[16] Chun-Zhi Z, Lei H, An-Ling Z, et al. MicroRNA-221 and microRNA-222 regulate gastric carcinoma cell proliferation and radioresistance by targeting PTEN[J]. BMC Cancer, 2010, 10: 367.
[17] Liu X, Yu J, Jiang L, et al. MicroRNA-222 regulates cell invasion by targeting matrix metalloproteinase 1 (MMP1) and manganese superoxide dismutase 2 (SOD2) in tongue squamous cell carcinoma cell lines[J]. Cancer Genomics Proteomics, 2009, 6(3): 131-139.
[18] Le Sage C, Nagel R, Egan DA, et al. Regulation of the p27(Kip1) tumor suppressor by miR-221 and miR-222 promotes cancer cell proliferation[J]. EMBO J, 2007, 26(15): 3699-3708.
[19] Visone R, Russol L, Pallante P, et al. MicroRNAs (miRNA)-221 and miRNA-222, both overexpressed in human thyroid papillary carcinomas, regulate p27Kip1 protein levels and cell cycle[J]. Endocr Relat Cancer, 2007, 14(3): 791-798.
[20] Zhang CZ, Zhang JX, Zhang AL, et al. MiR-221 and miR-222 target PUMA to induce cell survival in glioblastoma[J]. Mol Cancer, 2010, 9: 229.
[21] Firestein R, Blander G, Michan S, et al. The SIRT1 deacetylase suppresses intestinal tumorigenesis and colon cancer growth[J]. PLoS One, 2008, 3(4): e2020.
[22] Yuan J, Minter-Dykhouse K, Lou Z. A c-Myc-SIRT1 feedback loop regulates cell growth and transformation[J]. J Cell Biol, 2009, 185(2): 203-211.
[23] Dai Y, Ngo D, Forman LW, et al. Sirtuin 1 is required for antagonist-induced transcriptional repression of androgen-responsive genes by the androgen receptor[J]. Mol Endocrinol, 2007, 21(8): 1807-1821.
[24] Fu M, Liu M, Sauve AA, et al. Hormonal control of androgen receptor function through SIRT1[J]. Mol Cell Biol, 2006, 26(21):8122-8135.
[25] Lee H, Kim KR, Noh SJ, et al. Expression of DBC1 and SIRT1 is associated with poor prognosis for breast carcinoma[J]. Hum Pathol, 2011, 42(2): 204-213.
[26] Wang H, Liu H, Chen K, et al. SIRT1 promotes tumorigenesis of hepatocellular carcinoma through PI3K/PTEN/AKT signaling[J]. Oncol Rep, 2012, 28(1): 311-318.
[27] Huffman DM, Grizzle WE, Bamman MM, et al. SIRT1 is significantly elevated in mouse and human prostate cancer[J]. Cancer Res, 2007, 67(14): 6612-6618.
[28] Bradbury CA, Khanim FL, Hayden R, et al. Histone deacetylases in acute myeloid leukaemia show a distinctive pattern of expression that changes selectively in response to deacetylase inhibitors[J]. Leukemia, 2005, 19(10): 1751-1759.
[29] Stünkel W, Peh BK, Tan YC, et al. Function of the SIRT1 protein deacetylase in cancer[J]. Biotechnol J, 2007, 2(11): 1360-1368.
[30] Lim CS. SIRT1: tumor promoter or tumor suppressor[J]. Med Hypotheses, 2006, 7(2): 341-344.
[31] Ford J, Jiang M, Milner J. Cancer-specific functions of SIRT1 enable human epithelial cancer cell growth and survival[J]. Cancer Res, 2005, 65(22): 10457-10463.
[32] Pak S, Park S, Ryu J, et al. Preoperative factors predictive of posterolateral extracapsular extension after radical prostatectomy[J]. Korean J Urol, 2013, 54(12):824-829.