摘要
目的:探讨lncRNA MAGI2-AS3在神经母细胞瘤中的表达及对Fas/FasL通路的调控作用。方法:本研究选用2016年1月至2018年6月在本院接受治疗的23例神经母细胞瘤患者为研究对象,手术采集患者瘤组织和瘤旁正常组织进行qRT-PCR实验检测lncRNA MAGI2-AS3、Fas、FasL的mRNA的表达水平,并分析lncRNA MAGI2-AS3的表达与患者临床病理参数的关系;培养SH-SY5Y、SK-N-SH神经母细胞瘤细胞系,将其分为对照组(control)、LV-NC组、LV-MAGI2-AS3组,过表达lncRNA MAGI2-AS3,荧光显微镜检测慢病毒转染效率,qRT-PCR检测过表达后lncRNA MAGI2-AS3、Fas、FasL的mRNA的表达水平,CCK-8方法检测细胞增殖情况,流式细胞仪检测细胞凋亡情况。结果:神经母细胞瘤患者组织中lncRNA MAGI2-AS3、Fas、FasL mRNA表达水平均显著低于正常组织(P<0.05);与神经母细胞瘤患者临床病理参数比较结果显示lncRNA MAGI2-AS3与临床分期、组织分化程度有关(P<0.05);荧光显微镜观察SK-N-SH、SH-SY5Y细胞的转染效率,结果发现LV-MAGI2-AS3组显著高于LV-NC组(P<0.05),qRT-PCR检测lncRNA MAGI2-AS3 mRNA表达水平显著高于LV-NC组和control组(P<0.05);过表达LncRNA MAGI2-AS3后,两个细胞系中Fas、FasL mRNA表达水平显著高于LV-NC组和control(P<0.05);CCK-8检测结果显示,感染LV-MAGI2-AS3后,两个细胞系增殖率均显著低于LV-NC组和control组(P<0.05),LV-NC组和control组无显著变化;细胞凋亡检测结果显示,过表达LV-MAGI2-AS3z细胞凋亡率显著高于LV-NC组和control组(P<0.05),LV-NC组和control组凋亡率比较无显著差异。结论:lncRNA MAGI2-AS3在神经母细胞瘤组织和细胞中表达减少,且调控细胞凋亡通路中Fas、FasL的表达,其可能是通过调控Fas/FasL通路抑制神经母细胞瘤的生长。
Objective: To investigate the expression of lncRNA MAGI2-AS3 in neuroblastoma and its regulation on Fas/FasL pathway. Methods: 23 cases of neuroblastoma patients treated in our hospital from January 2016 to June 2018 were selected as the study subjects,the tumor tissues and adjacent normal tissues were collected,the expressions of lncRNA MAGI2-AS3,Fas and FasL mRNAs were detected by qRT-PCR,and the relationship between lncRNA MAGI2-AS3 expression with clinicopathological parameters was analyzed;SH-SY5Y and SK-N-SH neuroblastoma cell lines were cultured and divided into 3 groups:control group (control),LV-NC group and LV-MAGI2-AS3 group,made lncRNA MAGI2-AS3 over expression,the lentivirus transfection efficiency was detected by fluorescence microscopy,QRT-PCR detected the expression levels of mRNAs of lncRNA MAGI2-AS3,Fas and FasL after overexpression,CCK-8 assay was used to detect cell proliferation,and apoptosis was detected by flow cytometry. Results: The expression levels of lncRNA MAGI2-AS3,Fas and FasL mRNAs in neuroblastoma tissues were significantly lower than those in normal tissues ( P <0.05);the result compared with the clinicopathological parameters of neuroblastoma patients showed that lncRNA MAGI2-AS3 was associated with clinical stage and histological differentiation ( P <0.05);the transfection efficiencies of SK-N-SH and SH-SY5Y cells were observed by fluorescence microscope,the results showed that the LV-MAGI2-AS3 group was significantly higher than the LV-NC group ( P <0.05),qRT-PCR detection showed that the expression level of lncRNA MAGI2-AS3 mRNA was significantly higher than that in LV-NC group and control group ( P <0.05);after overexpression of lncRNA MAGI2-AS3,the expression levels of Fas and FasL mRNA in the two cell lines were significantly higher than those in LV-NC group and control group ( P <0.05);CCK-8 assay showed that,after infection with LV-MAGI2-AS3,the proliferation rates of the two cell lines were significantly lower than that of LV-NC group and control group ( P <0.05),there
作者
黄垂名
张小波
彭小旅
HUANG Chui-Ming;ZHANG Xiao-Bo;PENG Xiao-Lü(Hainan Provincial People′s Hospital,Haikou 570100,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2019年第16期1981-1986,共6页
Chinese Journal of Immunology
