组蛋白甲基化酶SMYD3在前列腺癌组织的表达及对mTOR通路的影响
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摘要
目的探讨组蛋白甲基化酶SMYD3在前列腺癌组织的表达水平及其对前列腺癌细胞生长的影响。方法免疫组化染色检测33例前列腺癌及癌旁组织的SMYD3表达水平;siRNA及SMYD3过表达质粒分别转染前列腺癌LNCa P和PC3细胞,检测其对前列腺癌细胞系生长的影响;Western blotting检测前列腺癌细胞中mTOR通路相关蛋白表达水平。结果 SMYD3在前列腺癌组织的表达水平明显高于癌旁组织,且SMYD3在胞核胞浆中均可检测到;转染SMYD3 siRNA后处理组LNCa P细胞在不同生长时期均少于对照组;而PC3细胞在转染SMYD3过表达质粒后细胞数则多于对照组。处理后,LNCa P细胞p-mTOR、p-p70S6K基因表达降低(P<0.05),而p-4EBP-1蛋白表达上升(P<0.05);PC3细胞呈现一致趋势。结论前列腺癌组织中SMYD3表达水平高于癌旁组织,且在胞核胞浆中均有表达;SMYD3表达可以促进前列腺癌细胞的生长;且可通过激活mTOR基因,参与mTOR通路的调节,影响前列腺癌细胞系的生长。
Objective To determine the expression of histone methyltransferase SMYD3 in prostate cancer tissues and to explore its role and mechanism in prostate cancer progression. Methods The expression of SMYD3 in 33 pairs of prostate cancer tissues and matched non-tumor tissues were detected with immunohistochemical staining. Prostate cancer cell line LNCa P and PC3 were transfected with siRNA and SMYD3 over-expressing vectors,respectively,to evaluate their growth. The expressions of proteins involved in mTOR pathway were evaluated with Western blotting. Results Prostate cancer had over-expressed SMYD3 compared with non-tumor normal tissues. The SMYD3 expression could be found in both nucleus and cytoplasm. SMYD3 abundance had a positive correlation with cell proliferation. The number of LNCa P cells was lower in SMYD3 siRNA group compared with the control group,and the number of PC3 cells was higher in SMYD3-PC vector group than in the control group. The depletion of the enzyme could down-regulate the expression of p-mTOR( P < 0. 05) and p-p70S6K( P < 0. 05),while the expression of p-4EBP-1( P < 0. 05) could be up-regulated; the trend of protein expression in mTOR pathway in PC3 was similar to that in LNCa P. Conclusion SMYD3 is over-expressed in prostate cancer tissues compared with non-tumor tissues,and SMYD3 can be expressed both in nucleus and cytoplasm. The expression of SMYD3 can activate the growth of prostate cancer cells by activating mTOR pathway.
Objective To determine the expression of histone methyltransferase SMYD3 in prostate cancer tissues and to explore its role and mechanism in prostate cancer progression. Methods The expression of SMYD3 in 33 pairs of prostate cancer tissues and matched non-tumor tissues were detected with immunohistochemical staining. Prostate cancer cell line LNCa P and PC3 were transfected with siRNA and SMYD3 over-expressing vectors,respectively,to evaluate their growth. The expressions of proteins involved in mTOR pathway were evaluated with Western blotting. Results Prostate cancer had over-expressed SMYD3 compared with non-tumor normal tissues. The SMYD3 expression could be found in both nucleus and cytoplasm. SMYD3 abundance had a positive correlation with cell proliferation. The number of LNCa P cells was lower in SMYD3 siRNA group compared with the control group,and the number of PC3 cells was higher in SMYD3-PC vector group than in the control group. The depletion of the enzyme could down-regulate the expression of p-mTOR( P < 0. 05) and p-p70S6K( P < 0. 05),while the expression of p-4EBP-1( P < 0. 05) could be up-regulated; the trend of protein expression in mTOR pathway in PC3 was similar to that in LNCa P. Conclusion SMYD3 is over-expressed in prostate cancer tissues compared with non-tumor tissues,and SMYD3 can be expressed both in nucleus and cytoplasm. The expression of SMYD3 can activate the growth of prostate cancer cells by activating mTOR pathway.
引文
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[12]Edlind MP,Hsieh AC.PI3K-AKT-m TOR signaling in prostate cancer progression and androgen deprivation therapy resistance[J].Asian J Androl,2014,16(3):378-386.
[13]Park H,Kim Y,Sul JW,et al.Synergistic anticancer efficacy of MEK inhibition and dual PI3K/m TOR inhibition in castration-resistant prostate cancer[J].Prostate,2015,75(15):1747-1759.
[14]Ali AK,Nandagopal N,Lee SH.IL-15-PI3K-AKTm TOR:a critical pathway in the life journey of natural killer cells[J].Front Immunol,2015,6:355.doi:10.3389/fimmu.2015.00355.e Collection 2015.
[15]Guertin DA,Sabatini DM.Defining the role of m TOR in cancer[J].Cancer Cell,2007,12(1):9-22.
[16]Yamamoto K,Ishida T,Nakano K,et al.SMYD3 interacts with HTLV-1 Tax and regulates subcellular localization of Tax[J].Cancer Sci,2011,102(1):260-266.
[17]Hayashi M,Deng L,Chen M,et al.Interaction of the hepatitis B virus X protein with the lysine methyltransferase SET and MYND domain-containing 3 induces activator protein 1 activation[J].Microbiol Immunol,2016,60(1):17-25.
[2]Lonergan PE,Tindall DJ.Androgen receptor signaling in prostate cancer development and progression[J].J Carcinog,2011,23.doi:10.4103/1477-3163.83937.
[3]Liu C,Wang C,Wang K,et al.SMYD3 as an oncogenic driver in prostate cancer by stimulation of androgen receptor transcription[J].J Natl Cancer Inst,2013,105(22):1719-1728.
[4]Nishio K,Sakai K,Togashi Y.PI3K and m TOR pathway and molecular targeted agents[J].Nihon Rinsho,2015,73(8):1315-1322.
[5]Liu Y,Bi T,Dai W,et al.Lupeol induces apoptosis and cell cycle arrest of human osteosarcoma cells through PI3K/AKT/m TOR Pathway[J].Technol Cancer Res Treat,2015.pii:1533034615609014.[Epub ahead of print]
[6]Follo MY,Manzoli L,Poli A,et al.PLC and PI3K/Akt/m TOR signalling in disease and cancer[J].Adv Biol Regul,2015,57:10-16.doi:10.1016/j.jbior.2014.10.004.Epub 2014 Nov 26.
[7]Guo Z,Hu X,Xing Z,et al.Baicalein inhibits prostate cancer cell growth and metastasis via the caveolin-1/AKT/m TOR pathway[J].Mol Cell Biochem,2015,406(1-2):111-119.
[8]Vieira FQ,Costa-Pinheiro P,Almeida-Rios D,et al.SMYD3 contributes to a more aggressive phenotype of prostate cancer and targets Cyclin D2 through H4K20me3[J].Oncotarget,2015,6(15):13644-13657.
[9]Bitting RL,Armstrong AJ.Targeting the PI3K/Akt/m TOR pathway in castration-resistant prostate cancer[J].Endocr Relat Cancer,2013,20(3):R83-R99.
[10]Jiang SJ,Wang S.Dual targeting of m TORC1 and m TORC2 by INK-128 potently inhibits human prostate cancer cell growth in vitro and in vivo[J].Tumour Biol,2015,36(10):8177-8184.
[11]李刚,闫若东,穆中一,等.m TOR对非激素依赖性前列腺癌的作用研究[J].肿瘤学杂志,2013,19(12):947-953.LI Gang,YAN Ruodong,MU Zhongyi,et al.Study on the effect of m TOR on androgen-independent prostate cancer[J].Journal of Chinese Oncology,2013,19(12):947-953.
[12]Edlind MP,Hsieh AC.PI3K-AKT-m TOR signaling in prostate cancer progression and androgen deprivation therapy resistance[J].Asian J Androl,2014,16(3):378-386.
[13]Park H,Kim Y,Sul JW,et al.Synergistic anticancer efficacy of MEK inhibition and dual PI3K/m TOR inhibition in castration-resistant prostate cancer[J].Prostate,2015,75(15):1747-1759.
[14]Ali AK,Nandagopal N,Lee SH.IL-15-PI3K-AKTm TOR:a critical pathway in the life journey of natural killer cells[J].Front Immunol,2015,6:355.doi:10.3389/fimmu.2015.00355.e Collection 2015.
[15]Guertin DA,Sabatini DM.Defining the role of m TOR in cancer[J].Cancer Cell,2007,12(1):9-22.
[16]Yamamoto K,Ishida T,Nakano K,et al.SMYD3 interacts with HTLV-1 Tax and regulates subcellular localization of Tax[J].Cancer Sci,2011,102(1):260-266.
[17]Hayashi M,Deng L,Chen M,et al.Interaction of the hepatitis B virus X protein with the lysine methyltransferase SET and MYND domain-containing 3 induces activator protein 1 activation[J].Microbiol Immunol,2016,60(1):17-25.