欢迎访问《解剖学报》官方网站！今天是
English
XAV939抑制人神经母细胞瘤细胞的增殖
Effect of XAV939 on proliferation inhibition of human neuroblastoma cell
目的 探讨小分子抑制剂XAV939对人神经母细胞瘤（NB）细胞系SH-SY5Y细胞的抗增殖和诱导凋亡的作用，及其可能机制。 方法 采用MTT法检测XAV939对SH-SY5Y细胞活力的抑制作用，并确定最佳作用浓度。然后采用Annexin V-FITC法检测XAV939处理后早期凋亡细胞百分比，Hoechst33342染色法观察凋亡细胞核形态，克隆形成实验检测细胞体外增殖能力的变化。Western blotting检测 Wnt/β-连环蛋白（β-catenin信号通路的关键蛋白和抗凋亡蛋白Bcl-2的变化。结果 MTT结果显示，1μmol/L XAV939作用24h后，SH-SY5Y 细胞增殖的抑制率有明显上升。XAV939处理后48h及72h，凋亡细胞百分比显著高于对照组，且细胞呈现出不同程度的凋亡形态。此外，XAV939可显著降低SH-SY5Y细胞的体外克隆形成率，降低Wnt/β-catenin信号通路关键蛋白β-catenin，Cyclin D1和c-Myc及抗凋亡蛋白Bcl-2的表达。 结论 XAV939可能部分通过抑制Wnt/β-catenin信号通路而抑制SH-SY5Y细胞的增殖。
Objective To study the effect of antiproliferative and apoptosis induction of small molecule inhibitor XAV939 on neuroblastoma (NB) cell line SH-SY5Y cells, and its possible mechanism. Methods The MTT assay was used to detect the inhibition effect of XAV939 on the viability of SH-SY5Y cells, and to determine the optimal concentration. Annexin V-FITC assay was used to detect the percentage of early apoptotic cells after XAV939 treatment. Hoechst33342 staining was used for observing the apoptotic nuclear morphology. Colony forming assay was applied for detecting the proliferation effect of XAV939 on SH-SY5Y cells. The expression of key proteins of Wnt/β-catenin pathway and anti-apoptotic protein Bcl-2 were detected by Western blotting. Results The MTT assay showed that the proliferation inhibition rate of SH-SY5Y cells increased significantly by 1μmol/L XAV939 for 24hours. The percentages of apoptotic cells were significantly higher than those in control groups after treated with XAV939 for 48hours and 72hours, and the cells showed varying degrees of apoptotic morphology. In addition, XAV939 significantly reduced the colony formation rate of SH-SY5Y cells in vitro. The results of Western blotting showed that XAV939 reduced the expression of key proteins of Wnt/β-catenin signaling pathway as well as anti-apoptotic protein Bcl-2. Conclusion XAV939 may inhibit the proliferation of SH-SY5Y cells in part by inhibiting the Wnt/β-catenin signaling pathway.
［1］Huang S M, Mishina YM, Liu S, et al. Tankyrase inhibition stabilizes axin and antagonizes Wnt sianaling［J］. Nature, 2009, 461(7264):614-620.
［2］Chen B, Dodge ME, Tang W, et al. Small molecule-mediated disruption of Wnt-dependent signaling in tissue regeneration and cancer［J］. Nat Chem Biol, 2009,5(2):100-107.［3］Zhi F, Gong G, Xu Y, et al. Activated β-catenin forces N2A cell-derived neurons back to tumor-like neuroblasts and positively correlates with a risk for human neuroblastoma［J］. Int J Biol Sci, 2012, 8(2):289-297.
［4］Jenkins D, Michael O, Mahendroo M, et al. Exon-specific northern analysis and rapid amplification of cdna ends (race) reveal that the proximal promoter ii (pii) is responsible for aromatase cytochrome p450 (cyp19) expression in human ovary［J］. Mol Cell Endocrinol, 1993, 97(1-2):R1-6.
［5］Liang Y, Zhong Z, Huang Y, et al. Stem-like cancer cells are inducible by increasing genomic instability in cancer cells［J］. J Biol Chem, 2010, 285(7):4931-4940.
［6］Maris JM, Matthay KK. Molecular biology of neuroblastoma［J］. J Clin Oncol, 1999, 17(7):2264-2279.
［7］Maris JM, Hogarty MD, Bagatell R, et al. Neuroblastoma［J］. Lancet, 2007, 369(9579):2106-2120.
［8］Sharp SE, Gelfand MJ, Shulkin B. Pediatrics: diagnosis of neuroblastoma［J］. Semin Nucl Med, 2011, 41(5):345-353.
［9］Bilir A, Erguven M, Yazihan N, et al. Enhancement of vinorelbine-induced cytotoxicity and apoptosis by clomipramine and lithium chloride in human neuroblastoma cancer cell line SH-SY5Y［J］. J Neurooncol, 2010, 100(3):385-395.
［10］Matthay KK, Villablanca JG, Seeger RC, et al. Treatment of high-risk neuroblastoma with intensive chemotherapy, radiotherapy, autologous bone marrow transplantation, and 13-cis-retinoic acid. Children’s Cancer Group［J］. N Engl J Med, 1999, 341(16):1165-1173.
［11］Pearson AD, Pinkerton CR, Lewis IJ, et al. High-dose rapid and standard induction chemotherapy for patients aged over 1 year with stage 4 neuroblastoma: a randomised trial［J］. Lancet Oncol, 2008, 9(3):247-256.
［12］Zage PE, Kletzel M, Murray K, et al. Outcomes of the POG 9340/9341/9342 trials for children with high-risk neuroblastoma: a report from the Children’s Oncology Group［J］.Pediatr Blood Cancer, 2008, 51(6):747-753.
［13］Lau L, Tai D, Weitzman S, et al. Factors influencing survival in children with recurrent neuroblastoma［J］. J Pediatr Hematol Oncol, 2004, 26(4):227-232.
［14］Laverdière C, Cheung NK, Kushner BH, et al. Long-term complications in survivors of advanced stage neuroblastoma［J］. Pediatr Blood Cancer, 2005, 45(3):324-332.
［15］Clevers H. Wnt/beta-catenin signaling in development and disease［J］. Cell, 2005, 127(3):469-480.
［16］Logan CY, Nusse R. The Wnt signaling pathway in development and disease［J］. Annu Rev Cell Dev Biol, 2004, 20(1):781-810.
［17］MacDonald BT, Tamai K, He X. Wnt/beta-catenin signaling: components, mechanisms, and diseases［J］. Dev Cell, 2009, 17(1):9-26.
［18］Barker N, Clevers H. Mining the Wnt pathway for cancer therapeutics［J］. Nat Rev Drug Discov, 2006, 5(12):997-1014.
［19］Dregalla RC, Zhou J, Idate RR, et al. Regulatory roles of tankyrase 1 at telomeres and in DNA repair: suppression of T-SCE and stabilization of DNA-PKcs［J］. Aging (Albany NY), 2010, 2(10):691-708.
［20］Damalas A, Ben-Ze’ev A, Simcha I, et al. Excess β-catenin promotes accumulation of transcriptionally active p53［J］. EMBO J, 1999,18(11):3054-3063.
［21］He TC, Sparks AB, Rago C, et al. Identification of c-MYC as a target of the APC pathway［J］. Science, 1998, 281(5382):1509-1512.
［22］Sun XF, Yin QW, Li XB, et al. The expression of anty-apoptosis protein BAG-1 in non-small cell lung cencerd and its relationship with cell apoptosis and expresson of Bcl-2 ［J］. Acta Anatomica Sinica, 2008, 29(4): 557-563. (in Chinese)
孙雪飞，尹秋伟，李希波，等.抗凋亡蛋白BAG.1在非小细胞肺癌组织中的表达及其与细胞凋亡、Bcl.2蛋白表达的关系［J］. 解剖学报，2008,39（4）：557-563.
［23］Ma WW, Adjei AA. Novel agents on the horizon for cancer therapy［J］. CA Cancer J Clin. 2010, 60(1):62.
镁金属相下成骨性骨髓细胞球生长动力学模型的建立及其迁移因素研究
/
| 〈 |
|
〉 |
