Effect of siRNA targeting HER-2/neu on proliferation of human glioma cells and its related gene expression

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Author:
ZHANG Heng(Department of Neurosurgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080,China)
WU Xin-jian(Department of Neurosurgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080,China)
YU Zhen-hua(Department of Neurosurgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080,China)
YANG Chao(Department of Neurosurgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080,China)
HUANG Zheng-song(Department of Neurosurgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080,China)
Journal Title:
CHINESE JOURNAL OF NEUROMEDICINE
Issue:
Volume 10, Issue 06, 2011
DOI:
10.3760/cma.j.issn.1671-8925.2011.06.004
Key Word:
HER-2/neu;Glioma;Proliferation;PKB

Abstract: Objective To investigate the effect of HER-2/neu siRNA on proliferation of human glioma cell lines U251MG and T98G which over-express HER-2/neu, and explore its mechanism.Methods Liposome-mediated HER-2/neu siRNA was transfected into human glioma cell lines U251MG and T98G;lipofectin group was established as controls. The mRNA and protein levels of HER-2/neu were detected by real-time PCR and Western blotting 3 d after the transfection. The proliferation of glioma cells was investigated using methyl thiazolyl tetrazolium (MTT) assay 3 and 4 d after the transfection. The effects of HER-2/neu siRNA on AKT/FOXO1 pathway and protein expression of p27 and Cyclin D1 were studied using Western blotting. Results HER-2/neu mRNA and protein expressions in the transfected U251MG cells were decreased to (28.833±4.174)% and (22.167±1.955)% while those in cells of the lipofectin group were (92.067±5.698)% and (96.100±1.682)%, respectively,with significant differences (P=0.000, 0.001). HER-2/neu mRNA and protein expressions of the transfected T98G cells were decreased to (28.067 ±6.165)% and (12.433 ±8.864)% while those in the untransfected cells were (96.000 ±5.110)% and (94.333 ±3.215)%, respectively, with significant differences (P=0.001, 0.008). Three d after the transfection, the rates of proliferation in the transfected T98G and U251MG cells were (58.467±5.561)% and (63.933±5.363)%, respectively;4 d after the transfection, the rates of proliferation in the transfected T98G and U251MG cells were (57.500±4.770)% and (60.167±3.253)%, respectively;an obvious decrease was noted as compared them with cells of the lipofectin group (P=0.020, 0.023, 0.021, 0.008). Cyclin Dl expression was decreased, while p27 protein expression was up-regulated in the transfected cells as compared with those in cells of the lipofectin group (P<0.05). Moreover, the levels of phosphorylated AKT and phosphorylated FOXO1 were decreased in the transfected cells as compared with those in cells of the lipofectin group (P<0.05).Conclusion The specific siRNA targeting HER-2/neu in human glioma cell lines U251MG and T98G could inhibit the cell proliferation, which might relate to the suppression of AKT/FOXO1 pathway and the regulation of expresion of thier downstream molecules such as p27 and Cyclin D1.

  • [1]Hirsch FR,Varella-Garcia M,Cappuzzo F.Predictive value of EGFR and HER2 overexpression in advanced non-small-cell lung cancer[J].Oncogene,2009,28(Suppl 1):S32-37.
  • [2]Shang J,Bi ZG,Ji HF,et al.Integrated assessment of potential value of erbB-2 amplification or expression status as novel therapy target in Chinese children and adolescents with osteosarcoma[J].Chin Med J(Engl),2009,122(13):1521-1524.
  • [3]周崧,殷晓璐,许雁萍,等.p16、FGFR-4及c-erbB-2的表达对垂体腺瘤生物学行为的意义[J].中华神经医学杂志,2008,7(8):784-787.
  • [4]李苏霞,张阳,徐建明,等.HER2、HER3蛋白表达与Gefitinib疗效的关系[J].实用医学杂志,2009,25(1):65-67.
  • [5]Andersson U,Schwartzbaum J,Wiklund F,et al.A comprehensive study of the association between the EGFR and ERBB2 genes and glioma risk[J].Acta Oncol,2010,49(6):767-775.
  • [6]Koka V,Potti A,Forseen SE,et al.Role of Her-2/neu overexpression and clinical determinants of early mortality in glioblastoma multiforme[J].Am J Clin Oncol,2003,26 (4):332-335.
  • [7]Choudhury A,Charo J,Parapuram SK,et al.Small interfering RNA (siRNA) inhibits the expression of the Her2/neu gene,upregulates HLA class I and induces apoptosis of Her2/neu positive tumor cell lines[J].Int J Cancer,2004,108(1):71-77.
  • [8]Mineo JF,Bordron A,Baroncini M,et al.Low HER2-expressing glioblastomas are more often secondary to anaplastic transformation of low-grade glioma[J].J Neurooncol,2007,85(3):281-287.
  • [9]Liu Z,Kokunai T,Tamaki N.Tamoxifen interacts with NEU/C-ERBB-2 receptor and inhibits growth of human malignant glioma cell lines[J].Kobe J Med Sci,2001,47(3):131-140.
  • [10]程俊,牛朝诗,潘琪.Skp2、P27kip1和PTEN在人脑胶质瘤中的表达及其相互关系的研究[J].中华神经医学杂志,2007,6(3):274-277.
  • [11]Gao L,Li F,Dong B,et al.Inhibition of STAT3 and ErbB2 suppresses tumor growth,enhances radiosensitivity,and induces mitochondria-dependent apoptosis in glioma cells[J].Int J Radiat Oncol Biol Phys,2010,77(4):1223-1231.
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