Effects of <i>EZH2</i>-shRNA on Proliferation and Invasion of Human Glioma Cells_West China Medical Journal

Authors：

 ZHOUZhang-ming ¹ , CHUSheng-hua ²

1. Department of Neurosurgery, Dujiangyan Medical Center, Dujiangyan, Sichuan 618300, P. R. China;
2. Department of Neurosurgery, No.3 People's Hospital Affiliated to School of Medicine, Shanghai Jiantong University, Shanghai 201999, P. R. China;

Corresponding author：

Keywords：

Glioma; EZH2; Proliferation; Invasion; RNA interference

DOI：

10.7507/1002-0179.20140608

Video：

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Objective To investigate the influence of EZH2 gene down-regulation by RNA interference on the proliferation and invasion of human glioma cell line U251. Methods The recombinant plasmid of small hairpin RNA targeting EZH2 gene was constructed, and transfected into gioma U251 cells by electroporation. The expression of EZH2 mRNA and protein in the cells was detected by using reverse transcriptase-polymerase chain reaction and Western blot respectively; the viability of cells was determined by using methyl thiazol tetrazo1ium assay; and the invasiveness of U251 cells was tested by Transwell cabin. Results The expression levels of EZH2 mRNA in U251 cells were detected in a significantly lower proportion in the EZH2-shRNA group (0.19±0.02) than that in the untransfected group (1.13±0.05) and the control-shRNA-GFP group (1.15±0.05). The expression levels of EZH2 protein in U251 cells were detected in a significantly lower proportion in the EZH2-shRNA group (0.20±0.02) than that in the untransfected group (1.03±0.03) and the control-shRNA-GFP group (0.97±0.06). The proliferation rates in EZH2-shRNA group were significantly decreased as compared with those in the untransfected group and control-shRNA-GFP group (24 hours after transfection:60.13%±3.15%, 100.00%±9.31%, 100.03%±9.35%; 48 hours after transfection:53.01%±3.14%, 100.00%±9.13%, 99.58%±9.27%; P<0.05) and Transwell cabin suggested that the invasiveness of U251 cells was significantly decreased (46.00±2.82, 60.67±5.71, 61.00±2.48; P<0.01). Conclusion EZH2-targeted RNA interference can reduce the proliferation and invasion of human glioma cells, which suggests that EZH2 shRNA may be a potential gene therapeutic target of human glioma.

Citation： ZHOUZhang-ming, CHUSheng-hua. Effects of EZH2-shRNA on Proliferation and Invasion of Human Glioma Cells. West China Medical Journal, 2014, 29(11): 2001-2005. doi: 10.7507/1002-0179.20140608 Copy

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5.	Reijm EA, Jansen MP, Ruigrok-Ritstier K, et al. Decreased expression of EZH2 is associated with upregulation of ER and favorable outcome to tamoxifen in advanced breast cancer[J]. Breast Cancer Res Treat, 2011, 125(2):387-394.
6.	Hinz S, Weikert S, Magheli A, et al. Expression profile of the polycomb group protein enhancer of Zeste homologue 2 and its prognostic relevance in renal cell carcinoma[J]. J Urol, 2009, 182(6):2920-2925.
7.	Mccabe MT, Creasy CL. EZH2 as a potential target in cancer therapy[J]. Epigenomics, 2014, 6(3):341-351.
8.	Debeb BG, Gong Y, Atkinson RL, et al. EZH2 expression correlates with locoregional recurrence after radiation in inflammatory breast cancer[J]. J Exp Clin Cancer Res, 2014, 33(1):58.
9.	Wee ZN, Li Z, Lee PL, et al. EZH2-mediated inactivation of IFN-γ-JAK-STAT1 signaling is an effective therapeutic target in MYC-driven prostate cancer[J]. Cell Rep, 2014, 8(1):204-216.
10.	Cavalli G. Molecular biology. EZH2 goes solo[J]. Science, 2012, 338(6113):1430-1431.
11.	刘细国, 袁先厚, 江普查, 等. STAT3和Cyclin D1在人脑胶质瘤中的表达及其意义[J]. 中华实验外科杂志, 2010, 27(10):1389-1391.
12.	Li XJ, Guo QH, Wang X, et al. LRP16 gene protects mouse insulinoma MIN6 cells against fatty acid-induced apoptosis through Akt/FoxO1 signaling[J]. Chin Med J (Engl), 2012, 125(10):1695-1702.
13.	Wallace GC, Haar CP, Vandergrift WA, et al. Multi-targeted DATS prevents tumor progression and promotes apoptosis in ectopic glioblastoma xenografts in SCID mice via HDAC inhibition[J]. J Neurooncol, 2013, 114(1):43-50.
14.	Qin L, Wang CZ, Fan HJ, et al. A dual-targeting liposome conjugated with transferrin and arginine-glycine-aspartic acid peptide for glioma-targeting therapy[J]. Oncol Lett, 2014, 8(5):2000-2006.
15.	Nagane M. Anti-angiogenic therapy for malignant glioma[J]. Gan To Kagaku Ryoho, 2014, 41(2):141-147.
16.	Sahebjam S, Mcnamara MG, Mason WP. Emerging biomarkers in anaplastic oligodendroglioma:implications for clinical investigation and patient management[J]. CNS Oncol, 2013, 2(4):351-358.

1. Yamaguchi H, Hung MC. Regulation and role of EZH2 in cancer[J]. Cancer Res Treat, 2014, 46(3):209-222.
2. Zheng S, Houseman EA, Morrison Z, et al. DNA hypermethylation profiles associated with glioma subtypes and EZH2 and IGFBP2 mRNA expression[J]. Neuro Oncol, 2011, 13(3):280-289.
3. Tao T, Liu D, Liu C, et al. Autoregulatory feedback loop of EZH2/miR-200c/E2F3 as a driving force for prostate cancer development[J]. Biochim Biophys Acta, 2014, 1839(9):858-865.
4. Bracken AP, Pasini D, Capra M, et al. EZH2 is downstream of the pRB-E2F pathway, essential for proliferation and amplified in cancer[J]. EMBO J, 2003, 22(20):5323-5335.
5. Reijm EA, Jansen MP, Ruigrok-Ritstier K, et al. Decreased expression of EZH2 is associated with upregulation of ER and favorable outcome to tamoxifen in advanced breast cancer[J]. Breast Cancer Res Treat, 2011, 125(2):387-394.
6. Hinz S, Weikert S, Magheli A, et al. Expression profile of the polycomb group protein enhancer of Zeste homologue 2 and its prognostic relevance in renal cell carcinoma[J]. J Urol, 2009, 182(6):2920-2925.
7. Mccabe MT, Creasy CL. EZH2 as a potential target in cancer therapy[J]. Epigenomics, 2014, 6(3):341-351.
8. Debeb BG, Gong Y, Atkinson RL, et al. EZH2 expression correlates with locoregional recurrence after radiation in inflammatory breast cancer[J]. J Exp Clin Cancer Res, 2014, 33(1):58.
9. Wee ZN, Li Z, Lee PL, et al. EZH2-mediated inactivation of IFN-γ-JAK-STAT1 signaling is an effective therapeutic target in MYC-driven prostate cancer[J]. Cell Rep, 2014, 8(1):204-216.
10. Cavalli G. Molecular biology. EZH2 goes solo[J]. Science, 2012, 338(6113):1430-1431.
11. 刘细国, 袁先厚, 江普查, 等. STAT3和Cyclin D1在人脑胶质瘤中的表达及其意义[J]. 中华实验外科杂志, 2010, 27(10):1389-1391.
12. Li XJ, Guo QH, Wang X, et al. LRP16 gene protects mouse insulinoma MIN6 cells against fatty acid-induced apoptosis through Akt/FoxO1 signaling[J]. Chin Med J (Engl), 2012, 125(10):1695-1702.
13. Wallace GC, Haar CP, Vandergrift WA, et al. Multi-targeted DATS prevents tumor progression and promotes apoptosis in ectopic glioblastoma xenografts in SCID mice via HDAC inhibition[J]. J Neurooncol, 2013, 114(1):43-50.
14. Qin L, Wang CZ, Fan HJ, et al. A dual-targeting liposome conjugated with transferrin and arginine-glycine-aspartic acid peptide for glioma-targeting therapy[J]. Oncol Lett, 2014, 8(5):2000-2006.
15. Nagane M. Anti-angiogenic therapy for malignant glioma[J]. Gan To Kagaku Ryoho, 2014, 41(2):141-147.
16. Sahebjam S, Mcnamara MG, Mason WP. Emerging biomarkers in anaplastic oligodendroglioma:implications for clinical investigation and patient management[J]. CNS Oncol, 2013, 2(4):351-358.

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