前列腺癌中Sp1通过PKM2途径促进细胞增殖代谢-东南大学学报医学版

前列腺癌中Sp1通过PKM2途径促进细胞增殖代谢

作者：刘大闯¹ 2 3 梁清¹ 陶陶² 3 黄烨清² 3 许斌² 3 陈恕求² 3 张治国¹ 董洋¹ 韩从辉¹ 陈明² 3

单位：1. 东南大学附属徐州市中心医院泌尿外科, 江苏徐州 221009;
2. 东南大学附属中大医院泌尿外科, 江苏南京 210009;
3. 东南大学医学院外科中心实验室, 江苏南京 210009

分类号：R737.25

出版年·卷·期（页码）：2015·34·第三期（337-341）

摘要：

目的:确定前列腺癌中Sp1通过PKM2途径对前列腺癌细胞的增殖和代谢的作用.方法:前列腺癌细胞株DU145与PC3体外转染Sp1 siRNA与阴性对照无意义核苷酸序列(NC组),采用葡萄糖检测试剂盒与乳酸检测试剂盒检测转染后有氧糖酵解变化;CCK-8实验检测转染后的细胞增殖; qRT-PCR检测转染后PKM2表达;Western Blotting检测转染后Sp1与PKM2表达.结果:DU145与PC3转染Sp1 siRNA后与NC组比较,剩余葡萄糖显著偏高,乳酸产生显著下降;CCK8实验表明抑制Sp1后能显著抑制前列腺癌细胞的增殖能力;qRT-PCR实验显示抑制Sp1后明显抑制PKM2的表达;Western Blotting显示转染的Sp1 siRNA对Sp1具有较高的沉默效率,且PKM2的表达也降低.结论:Sp1在前列腺癌细胞株DU145与PC3中可通过直接作用于PKM2促进细胞代谢.

Objective: To determine wether the Sp1 could accelerate the metabolism of prostate cancer cells through PKM2.Methods: DU145 and PC3 cell lines of prostate cancer were transfected with Sp1 siRNA or NC. Aerobic glycolysis was detected by the glucose and lactic acid detection kit after transfection. Cell proliferation was detected by CCK-8 assay. PKM2 mRNA expression was detected by qRT-PCR. Sp1or PKM2 expression was tested by Western Blotting. Results: The remaining glucose level was higher in Sp1 siRNA group than in NC group, the lactic acid level declined significantly after Sp1 siRNA transfection. CCK-8 assay revealed over-expression of Sp1 siRNA significantly inhibited proliferation. qRT-PCR showed that the inhibition of Sp1 could significantly inhibit the expression of PKM2. Western Blotting showed that Sp1 had high efficiency of silence, PKM2 expression reduced significantly after Sp1 siRNA reansfection.Conclusion: Sp1 acts directly on PKM2 to promote cell metabolism in DU145 and PC3 cell lines of prostate cancer.

参考文献：

[1] BLACK A R,BLACK J D,AZIZKHAN-CLIFFORD J.Sp1 and krüppel-like factor family of transcription factors in cell growth regulation and cancer[J].J Cell Physiol,2001,188(2):143-160.
[2] MAZUREK S,BOSCHEK C B,HUGO F,et al.Pyuvate kinase type M2 and its role in tumor growth and spreading[J].Semin Cancer Biol,2005,15(4):300-308.
[3] WU X,ZHOU Y,ZHANG K,et al.Isoform-specific interactionof pyruvate kinase with hepatitis C virus NS5B[J].FEBS Lett,2008,582(15):2155-2160.
[4] LEE J,KIM H K,HAN Y M,et al.Pyruvate kinase isozyme type M2(PKM2) interacts and cooperates with Oct-4 in regulating transcription[J].Int J Biochem Cell Bid,2008,40(5):1043-1054.
[5] DISCHER D J,BISHOPRIC N H,WU X,et al.Hypoxia regulates beta-enolase and pyruvate kinase-M promoters by modulating Sp1/Sp3 binding to a conserved GC element[J].J Biol Chem,1998,273(40):26087-26093.
[6] FELDMAN B J,FELDMAN D.The development of androgen independent prostate cancer[J].Nat Rev Cancer,2001,1(1):34-45.
[7] BEAVER L M,BUCHANAN A,SOKOLOWSKI E I,et al.Transcriptome analysis reveals a dynamic and differential transcriptional response to sulforaphane in normal andprostate cancer cells and suggests a role for Sp1 in chemoprevention[J].Mol Nutr Food Res,2014,58(10):2001-2013.
[8] LI Y,VONGSANGNAK W,CHEN L,et al.Integrative analysis reveals disease-associated genes and biomarkers for prostate cancer progression[J].BMC Med Genomics, 2014,7(Suppl 1):S3.
[9] JIANG J,JIA P,ZHAO Z,et al.Key regulators in prostate cancer identified by co-expression module analysis[J].BMC Genomics,2014,15:1015.
[10] 王琪炜,陈宝俊,强倩,等.人转录因子sp1在大肠杆菌中的表达与体外纯化[J].东南大学学报:医学版,2014,33(1):1-4.
[11] CUI Y,NADIMINTY N,LIU C,et al.Upregulation of glucose metabolism by NF-κB2/p52 mediates enzalutamide resistance in castration-resistantprostate cancer cells[J].Endocr Relat Cancer, 2014,21(3):435-442.
[12] LV L,LI D,ZHAO D,et al.Acetylation targets the M2 isoform of pyruvate kinase for degradation through chaperone-mediated autophagy and promotes tumor growth[J].Mol Cell, 2011,42(6):719-730.
[13] WONG N,YAN J,OJO D,et al.Changes in PKM2 associate with prostate cancer progression[J].Cancer Invest, 2014,32(7):330-338.
[14] 吕磊,袁敬东,操作亮,等.miR-124 通过靶向调控PKM2 基因的表达抑制前列腺癌PC3 细胞的增殖[J].中华男科学杂志,2014,20(6):495-499.
[15] 吕磊,汪良,蒋国松,等.沉默PKM2 增强藤黄酸诱导人前列腺癌PC3 细胞凋亡的敏感性[J].中华男科学杂志,2013,19(2):102-106.

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