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PED/PEA-15在前列腺癌中的表达及对前列腺癌细胞凋亡的影响 被引量：2

Expression of PED/PEA-15 in Prostate Cancer and Its Effect on Apoptosis of Prostate Cancer Cell
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摘要 目的通过构建PED/PEA-15特异的siRNA载体,探讨PED/PEA-15对前列腺癌细胞(PC-3)凋亡的影响。方法应用免疫组化SP法检测前列腺癌和正常前列腺组织中PED/PEA-15的表达;用RT-PCR法检测PC-3细胞中PED/PEA-15的表达。体外合成PED/PEA-15特异性siRNA序列,并克隆入真核表达载体psiRNA-hH1neo。以脂质体法转染psiRNA-PED/PEA-15至PC-3细胞中,以RT-PCR和Western blot法检测psiRNA-PED/PEA-15对PED/PEA-15表达的影响;用MTT和流式细胞术检测其对PC-3细胞凋亡的影响。结果免疫组化和RT-PCR均显示PED/PEA-15在前列腺癌中高表达;正常前列腺组织没有或只有很弱的表达。酶切和DNA测序证实合成的siRNA基因序列正确,并已被准确克隆入psiRNA-hH1neo载体。psiRNA-PED/PEA-15可特异性抑制PC-3细胞中PED/PEA-15的表达。转染psiRNA-PED/PEA-15的PC-3细胞凋亡显著增加(P<0.05)。结论PED/PEA-15可阻抑前列腺癌细胞凋亡,其表达对前列腺癌的发展有重要作用。 Objective To explore the effect of PED/PEA-15 on prostate cancer cell （PC-3） apoptosis by constructing PED/ PEA-15 specific siRNA vector. Methods The expression of PED/PEA-15 in prostate cancer cell （PC-3） and prostate tissue was respectively assayed by means of RT-PCR and immunohistochemistry techniques. The in vitro synthesized PED/PEA-15 specific siRNA sequence was cloned into psiRNA-hHlneo vector. The constructed psiRNA-PED/PEA-15 was transiently transfected into PC-3 cells by liposome. The inhibitory effect of psiRNA-PED/PEA-15 on the expression of PED/PEA-15 was detected by using RT-PCR and Western blot, and those on apoptosis of cancer cells by flow cytomtry and MTT techniques. Results Immunohistochemistry and RT-PCR revealed that PED/ PEA-15 was highly expressed in PC-3 ceils, while there was no or weak expression of PED/PEA-15 in normal prostate tissues. Enzyme digestion analysis and DNA sequencing confirmed that the PED/PEA-15 ,specific ,siRNA expression vector was successfully constructed and cloned into psiRNA-hHqneo vector. PsiRNA-PED/PEA-15 could specifically inhibit the expression of PED/PEA-15 in PC-3 cells. The apoptosis of PC-3 cells transfected with psiRNA-PED/PEA-15 was significantly increased （P〈0.05）. Conclusion PED/PEA-15 can inhibit the apoptosis of PC-3 cells, and its expression might be involved in prostate cancer development.
作者 胡晓勇 陈晓春 朱朝辉 曾甫清 鲁功成
机构地区 华中科技大学同济医学院附属协和医院泌尿外科
出处 《华中科技大学学报（医学版）》 CAS CSCD 北大核心 2006年第1期18-22,共5页 Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金 国家自然科学基金资助项目(No.30070756)
关键词 PED/PEA-15蛋白 前列腺癌 细胞凋亡 phosphoprotein enriched in diabetes/phosphoprotcin enriched in astrocytes prostate cancer apoptosis
分类号 R273 [医药卫生—中西医结合] Q55 [生物学—生物化学]
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参考文献11

  • 1CONDORELLI G,VIGLIOTTA G,IAVARONE C,et al.PED/PEA-15 gene controls glucose transport and is over-expressed in type 2 diabetes mellitus[J].EMBO J,1998,17(14):3858-3866.
  • 2GAUMONT-LECLERC M F,MUKHOPADHYAY U K,GOUMARD S,et al.PEA-15 is inhibited by adenovirus E1A and plays a role in ERK nuclear export and Ras-induced senescence[J].J Biol Chem,2004,279(45):46802-46809.
  • 3ALESSANDRA T,FRANCESCA F,MARIA A,et al.Omi/HtrA2 promotes cell death by binding and degrading the anti-apoptotic protein PED/PEA-15[J].J Boil Chem,2004,279(45):46566-46572.
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同被引文献7

  • 1胡晓勇,陈晓春,陈朝晖,曾甫清,鲁功成.Omi／HtrA2对前列腺癌细胞的促凋亡作用[J].中华实验外科杂志,2006,23(3):344-347. 被引量：7
  • 2Kim E H,Kim S U,Shin D Y,et al.Roseovitine sensitizes glioma cells to TRAIL-mediated apoptosis by downregulation of survivin and XIA P[J].Oncogene,2004,23 (2):446-456.
  • 3Shiraki K,Sugiraoto K,Yamanaka Y,et al.Overexpression of Xlinked inhibitor of apoptosis in human hepatocellular carcinoma[J].Int J Mol Med,2003,12(5):705-708.
  • 4Gaumont Leclerc M F,Mukhopadhyay U K,Goumard S,et al.PEA-15 is inhibited by adenovirus E1A and plays a role in ERK nuclear export and ras-induced senescence[J].J Biol Chem,2004,279 (45):46802-46809.
  • 5Hao C,Beguinot F,Condorelli G,et al.Induction and intracellular regulation of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) mediated apotosis in human malignant glioma cells[J].Cancer Res,2001,61 (3):1162-1170.
  • 6Stassi G,Garofalo M,Zerilli M,et al.PED mediates AKT-dependent chemoresistance in human breast cancer cells[J].Cancer Res,2005,65(15):6668-6675.
  • 7Trencia A, Fiory F, Alessandro M, et al. Omi/HtrA2 promotes cell death by binding and degrading the anti-apoptotic protein PED/PEA-15. J Boil Chem, 2004, 279: 46566-46572.

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华中科技大学学报（医学版）
2006年 第1期

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