MYCN基因表达抑制促TRAIL诱导神经母细胞瘤细胞凋亡的研究

摘要

Objective To investigate whether MYCN gene silence could enhance TRAIL induced apoptosis in MYCN amplified neuroblastoma cell line SK-N-BE ( 2) . Methods NB cells line SK-N-BE ( 2) were grouped into non-silencing siRNA group, TRAIL group, MYCN siRNA group and MYCN siRNA + TRAIL group. The MYCN siRNA was transfected to NB cell lines SK-N-BE ( 2) for gene silence. Western-blot was used to detect the expression of Bcl-2, Bcl-xL, Bid, DR4 and DR5; ELISA was used to detect the apoptosis of each group. Results Western-blot revealed that the MYCN siRNA decreased the expression of MYCN gene significantly ( P <0. 05) . After MYCN gene silence, the expression of DR5 and Bid were increased ( P < 0. 05 ) , the expression of Bcl-2 and Bcl-xL were decreased ( P < 0.05) , while the expression of DR4 did not change ( P > 0. 05) . MYCN gene silence could enhance TRAIL-induced apoptosis in MYCN amplified NB cell line SK-N-BE ( 2) ( P <0. 05) . Conclusions MYCN gene silence could enhance TRAIL induced apoptosis in MYCN amplified NB cell line SK-N-BE (2) by regulating the expression of Bel protein family and DR5.%目的 神经母细胞瘤(NB)为小儿常见恶性肿瘤.MYCN基因高扩增NB细胞对TRAIL诱导凋亡多有抵抗.我们以MYCN基因高扩增NB细胞株为研究对象,抑制其MYCN基因的表达,探讨其对TRAIL诱导凋亡有无增强作用.方法 构建MYCN siRNA,转染NB细胞株SK-N-BE(2),抑制MYCN基因表达后,Western-blot法检测SK-N-BE(2)细胞Bcl-2、Bcl-xL、Bid、DR4、DR5表达的变化,ELISA法检测细胞凋亡.结果 转染MYCN siRNA的SK-N-BE(2)细胞中MY-CN基因表达显著降低(P＜0.05).MYCN基因表达抑制后,sK-N-BE(2)细胞DR5、Bid表达增高(P＜0.05),Bcl-2、Bcl-xL表达降低(P＜0.05),DR4表达无变化(P＞0.05),TRAIL诱导细胞凋亡显著增强(P＜0.05).结论 MYCN基因表达抑制后,可调控Bcl家族中相关蛋白、DR5的表达,促进TRAIL诱导NB细胞凋亡.