Vitali Roberta, Mancini Camillo, Cesi Vincenzo, Tanno Barbara, Mancuso Mariateresa, Bossi Gianluca, Zhang Ying, Martinez Robert V, Calabretta Bruno, Dominici Carlo, Raschellà Giuseppe
Section of Toxicology and Biomedical Sciences, ENEA Research Center Casaccia, Rome, Italy.
Clin Cancer Res. 2008 Jul 15;14(14):4622-30. doi: 10.1158/1078-0432.CCR-07-5210.
We assessed the relevance of Slug (SNAI2) for apoptosis resistance and invasion potential of neuroblastoma cells in vitro and in vivo.
We evaluated the effect of imatinib mesylate on invasion and analyzed the genes modulated by imatinib mesylate treatment in neuroblastoma cells. Slug expression, inhibited by imatinib mesylate treatment, was knocked down in neuroblastoma cells by RNA interference, and the effects on invasion and apoptosis were evaluated in vitro. A pseudometastatic model of neuroblastoma in severe combined immunodeficient mice was used to assess the effects of Slug silencing alone or in combination with imatinib mesylate treatment on metastasis development.
Microarray analysis revealed that several genes, including Slug, were down-regulated by imatinib mesylate. Slug expression was detectable in 8 of 10 human neuroblastoma cell lines. Two Slug-expressing cell lines were infected with a vector encoding a microRNA to Slug mRNA. Infected cells with reduced levels of Slug were tested for the expression of apoptosis-related genes (p53, Bax, and Bcl-2) identified previously as Slug targets. Bcl-2 was down-regulated in Slug-interfered cells. Slug down-regulation increased sensitivity to apoptosis induced by imatinib mesylate, etoposide, or doxorubicin. Invasion of Slug-silenced cells was reduced in vitro. Animals injected with Slug-silenced cells had fewer tumors than controls and the inhibition of tumor growth was even higher in animals treated with imatinib mesylate.
Slug down-regulation facilitates apoptosis induced by proapoptotic drugs in neuroblastoma cells and decreases their invasion capability in vitro and in vivo. Slug inhibition, possibly combined with imatinib mesylate, may represent a novel strategy for treatment of metastatic neuroblastoma.
我们评估了Slug（SNAI2）在体外和体内对神经母细胞瘤细胞抗凋亡能力和侵袭潜能的相关性。
我们评估了甲磺酸伊马替尼对侵袭的影响，并分析了甲磺酸伊马替尼处理神经母细胞瘤细胞后所调节的基因。通过RNA干扰在神经母细胞瘤细胞中敲低受甲磺酸伊马替尼处理抑制的Slug表达，并在体外评估其对侵袭和凋亡的影响。利用严重联合免疫缺陷小鼠的神经母细胞瘤假转移模型，评估单独沉默Slug或与甲磺酸伊马替尼联合处理对转移发展的影响。
微阵列分析显示，包括Slug在内的几个基因被甲磺酸伊马替尼下调。在10个人类神经母细胞瘤细胞系中的8个中可检测到Slug表达。用编码针对Slug mRNA的微小RNA的载体感染两个表达Slug的细胞系。对Slug水平降低的感染细胞检测先前确定为Slug靶标的凋亡相关基因（p53、Bax和Bcl-2）的表达。Bcl-2在Slug干扰的细胞中下调。Slug下调增加了对甲磺酸伊马替尼、依托泊苷或阿霉素诱导的凋亡的敏感性。体外Slug沉默细胞的侵袭减少。注射了Slug沉默细胞的动物比对照组的肿瘤更少，在用甲磺酸伊马替尼处理的动物中肿瘤生长的抑制甚至更高。
Slug下调促进神经母细胞瘤细胞中促凋亡药物诱导的凋亡，并降低其在体外和体内侵袭能力。抑制Slug，可能与甲磺酸伊马替尼联合使用，可能代表一种治疗转移性神经母细胞瘤的新策略。
