Hematology, Oncology and Stem Cell Transplantation Research Center, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran.
Neurochem Res. 2012 Sep;37(9):2053-63. doi: 10.1007/s11064-012-0827-9. Epub 2012 Jun 21.
Neuroblastoma is the most common solid tumor in children. Current therapy modalities have resulted in little amelioration in the cure rate of neuroblsatoma and therefore, outlining biologically based therapies for neuroblastoma remains of main priority. This study was carried out to appraise the impeding effects of silibinin, a potent anti-cancer agent, on two different neuroblastoma cell lines, stromal SK-N-MC and neuroblastic SK-N-BE(2) cells. The microculture tetrazolium assay, gelatin zymography, colony formation assay, cell cycle distribution survey, apoptosis assay, and quantitative real-time reverse transcription-PCR were applied to evaluate the effects of silibinin on metabolic activity, gelatinolytic activity of MMP-2 and MMP-9, surviving potential, cell cycle, apoptosis, and expression pattern of the genes involved in cell survival and invasion of the two neuroblastoma cell lines. Treatment for 48 h inhibited metabolic activity and clonogenic potential of SK-N-MC cells in a dose-dependent manner. Silibinin also inhibited transcriptional levels of MMP-2, MMP-9, and uPAR, as markers of cell invasion, in SK-N-MC cells. Higher concentration of silibinin (75, 100 μM) suppressed enzymatic activity of MMP-2 in this cell line. No change in apoptosis and cell cycle was observed in neither of the cells after treatment with silibinin. On the other hand, silibinin highly decreased mRNA expression of Akt, and NF-κB1 and its regulators, IKK1 and IKK2 in SK-N-MC cell line. Comparison of transcriptional expression of Akt, and NF-κB1 in untreated stromal and neuroblastic cell lines shows that their basal transcriptional levels are much higher in SK-N-BE(2) cell line than that in SK-N-MC cells. It seems that SK-N-BE(2) cell line probably resists to silibinin through higher expression of Akt and probably NF-κB1. Collectively, our results demonstrated that silibinin highly inhibits the proliferative potentials of SK-N-MC cell line, whilst it had less inhibitory effect on SK-N-BE(2) cell line. Our results suggest that suppression of SK-N-MC cell line by silibinin may be through inhibition of Akt-mediated NF-κB1.
神经母细胞瘤是儿童中最常见的实体肿瘤。目前的治疗方法并没有显著提高神经母细胞瘤的治愈率，因此，制定基于生物学的神经母细胞瘤治疗方法仍然是当务之急。本研究旨在评估具有强大抗癌作用的水飞蓟素对两种不同的神经母细胞瘤细胞系（基质 SK-N-MC 和神经母细胞瘤 SK-N-BE(2)细胞）的抑制作用。微培养四唑盐法、明胶酶谱法、集落形成实验、细胞周期分布检测、凋亡检测和实时定量逆转录-PCR 用于评估水飞蓟素对代谢活性、MMP-2 和 MMP-9 的明胶酶活性、存活能力、细胞周期、凋亡以及两种神经母细胞瘤细胞系中与细胞存活和侵袭相关的基因表达模式的影响。48 小时的处理以剂量依赖的方式抑制 SK-N-MC 细胞的代谢活性和集落形成能力。水飞蓟素还抑制了 SK-N-MC 细胞中细胞侵袭标志物 MMP-2、MMP-9 和 uPAR 的转录水平。较高浓度的水飞蓟素（75、100μM）抑制了该细胞系中 MMP-2 的酶活性。水飞蓟素处理后，两种细胞的凋亡和细胞周期均无变化。另一方面，水飞蓟素在 SK-N-MC 细胞系中高度降低了 Akt 和 NF-κB1 及其调节因子 IKK1 和 IKK2 的 mRNA 表达。比较未经处理的基质和神经母细胞瘤细胞系中 Akt 和 NF-κB1 的转录表达表明，SK-N-BE(2)细胞系的基础转录水平远高于 SK-N-MC 细胞系。似乎 SK-N-BE(2)细胞系可能通过更高表达 Akt 并可能通过 NF-κB1 来抵抗水飞蓟素。总的来说，我们的结果表明，水飞蓟素强烈抑制 SK-N-MC 细胞系的增殖潜力，而对 SK-N-BE(2)细胞系的抑制作用较小。我们的结果表明，水飞蓟素对 SK-N-MC 细胞系的抑制作用可能是通过抑制 Akt 介导的 NF-κB1。
