Small interfering RNA -mediated cathepsin D knock-down impairing biological behavior of glioma cells

( views:348, downloads:0 )
Author:
ZHOU Jin-qiao(Department of Neurosurgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China)
WANG Jing-tao(Department of Neurosurgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China)
LIU Qiu-hong(Department of Neurosurgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China)
GUO Xin-bing(Department of Neurosurgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China)
SONG Lai-jun(Department of Neurosurgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China)
Journal Title:
Chinese Journal of Neuromedicine
Issue:
Volume 11, Issue 09, 2012
DOI:
10.3760/cma.j.issn.1671-8925.2012.09.001
Key Word:
Glioma;Small interfering RNA;Cathepsin D

Abstract: Objective To explore the effect of small interfering RNA (siRNA)-mediated cathepsin D (CD) knock-down on biological behavior of U87 cells. Methods CD-siRNA,scramble siRNA and control siRNA mediated by lipofectamin were introduced into the U87 cells,respectively.The silencing effect of CD was verified by RT-PCR and Western blotting in terms of CD mRNA and protein expressions. The cell viability of scramble and CD siRNA treated U87 cells was measured by MTT on 1-5 d after transfection. The invasive potential of scramble and CD siRNA treated U87 cells was measured by Transwell coated with Matrigel 48 h after transfection. Results Forty-eight h after transfection, mRNA and protein expressions of CD in CD siRNA treated U87 cells were remarkably reduced as compared with those in the scramble siRNA and control siRNA treated U87 cells (P<0.05).The absorbance (A value) of CD siRNA treated U87 cells was significantly lower than that in the scramble siRNA treated U87 cells 2,3,4 and 5 d after transfection (P<0.05); and the invasive potential of CD siRNA treated U87 cells was suppressed significantly as compared with that in the scramble siRNA treated U87 cells (P<0.05). Conclusion CD has the potential to be a promising therapeutic target.

  • [1]Chen ZG,Broddus WC,Zhu C,et al.Current status of genetherapy for gliomas[J].Acad J Sec Mil Med Univ,2004,25(1):96-100.
  • [2]曾义,游潮.RNAi技术及其在胶质瘤治疗中的研究进展[J].中华肿瘤防治杂志,2008,15(8):628-631.
  • [3]Brummelkamp TR.A system for stable expression of short in-terfering RNAs in mammalian cells[J]. Science,2002,296(5567):550-553.
  • [4]刘刚,陈谦学,吴立权,等.整合素β3和组织蛋白酶D在人脑胶质瘤中的表达及意义[J].中国临床神经外科杂志,2006,11(1):26-28.
  • [5]邢变枝,毛善平.Cathepsin D在脑星形胶质细胞瘤中的表达及对预后的评估效应[J].中国临床康复,2005,9(10):120-121.
  • [6]徐如祥.神经胶质瘤的基因治疗策略现状和未来[J].中华神经医学杂志.2003,2(6):401-403.
  • [7]Barthell E,Mylonas I,Shabani N.et al.Immunohistochemical visualisation of cathepsin-D expression in breast cancer[J].Anticancer Res,2007,27(4A):2035-2039.
  • [8]Minarowska A,Minarowski L,Karwowska A,et al.Regulatory role of cathepsin D in apoptosis[J].Folia Histochem Cytobiol,2007,45(3):159-163.
  • [9]Khalkhali-Ellis Z,Hendrix MJ.Elucidating the function of secreted maspin: inhibiting cathepsin D-mediated matrix degradation[J].Cancer Res,2007,67(8):3535-3539.
  • [10]Hood JD,Cheresh DA.Role of integrins in cell invasion and migration[J].Nat Rev Cancer,2002,2(2):91-100.
  • [11]Benes P,Vetvicka V,Fusek Met,et al.Cathepsin D many functions of one aspartic protease[J]. Crit Rev Oncol Hematol,2008,68(1):12-28.
  • [12]Nomura T,Katunuma N.Involvement of cathepsins in the invasion,metastasis and proliferation of cancer cells[J].J Med Invest,2005,52(1-2):1-9.
  • [13]Leto G,Tumminello FM,Crescimanno M,et al.Cathepsin D expression levels in nongynecological solid tumors: clinical and therapeutic implications[J].Clinical and Experimental metastasis,2004,21:91-106.
  • [14]Fan QW,Weiss WA.RNA interference against a glioma-derived allele of EGFR induces blockade at G2M[J].Oncogene,2005,24(5):829-837.
  • [15]杨翔云,赖小刚,张勇,等.siRNA干扰C1C-2表达对人胶质瘤U-87细胞增殖的抑制作用[J].癌症,2006,25(7):805-810.
  • [16]赵澎,胡微,张亚卓,等.RNA干扰技术逆转神经胶质瘤细胞多药耐药性[J].中华肿瘤杂志,2006,28(3):183-187.
WanfangData CO.,Ltd All Rights Reserved
About WanfangData | Contact US
Healthcare Department, Fuxing Road NO.15, Haidian District Beijing, 100038 P.R.China
Tel:+86-010-58882616 Fax:+86-010-58882615 Email:yiyao@wanfangdata.com.cn