中华小儿外科杂志
中華小兒外科雜誌
중화소인외과잡지
CHINESE JOURNAL OF PEDIATRIC SURGERY
2014年
5期
338-342
,共5页
潘敏%陈鑫%张虹%韩芦芦%鹿洪亭%郝希伟%董蒨
潘敏%陳鑫%張虹%韓蘆蘆%鹿洪亭%郝希偉%董蒨
반민%진흠%장홍%한호호%록홍정%학희위%동천
微RNAs%神经母细胞瘤%肿瘤转移
微RNAs%神經母細胞瘤%腫瘤轉移
미RNAs%신경모세포류%종류전이
microRNAs%Neuroblastoma%Neoplasm metastasis
目的 检测miR-34a在原发性和转移性神经母细胞瘤组织中的表达水平,并探讨miR-34a对神经母细胞瘤细胞生长、侵袭和迁移的影响.方法 收集18例神经母细胞瘤患儿(神经母细胞瘤原发部位以及转移部位),利用实时定量PCR检测miR-34a在原发性和转移性神经母细胞瘤组织中的表达水平.在神经母细胞瘤细胞中瞬时转染miR-34a mimics,利用平板克隆形成实验、MTT、侵袭实验和体外划痕实验检测miR-34a对细胞生长、侵袭和迁移的影响.最后利用Western印迹检测miR-34a对生长以及侵袭迁移相关蛋白表达水平的影响.结果 与原发性神经母细胞瘤组织相比,miR-34a在转移性神经母细胞瘤组织中表达降低.与对照细胞组相比,转染miR-34a mimics的细胞克隆数目降低(SH-SY5Y:24±3.07比43±6.29和IMR-32:17±2.36比37±5.41),细胞活性下降,并且细胞侵袭和迁移能力也降低(差异具有统计学意义).Western结果显示miR-34a抑制CD44、CDK4、CCND1和CCNE2的蛋白水平.结论 miR-34a分别通过降低CD44、CDK4、CCND1和CCNE2的表达水平,而抑制神经母细胞瘤细胞的生长、侵袭和迁移,发挥着肿瘤抑癌基因的作用,提示miR-34a可能作为神经母细胞瘤分子治疗的靶点.
目的 檢測miR-34a在原髮性和轉移性神經母細胞瘤組織中的錶達水平,併探討miR-34a對神經母細胞瘤細胞生長、侵襲和遷移的影響.方法 收集18例神經母細胞瘤患兒(神經母細胞瘤原髮部位以及轉移部位),利用實時定量PCR檢測miR-34a在原髮性和轉移性神經母細胞瘤組織中的錶達水平.在神經母細胞瘤細胞中瞬時轉染miR-34a mimics,利用平闆剋隆形成實驗、MTT、侵襲實驗和體外劃痕實驗檢測miR-34a對細胞生長、侵襲和遷移的影響.最後利用Western印跡檢測miR-34a對生長以及侵襲遷移相關蛋白錶達水平的影響.結果 與原髮性神經母細胞瘤組織相比,miR-34a在轉移性神經母細胞瘤組織中錶達降低.與對照細胞組相比,轉染miR-34a mimics的細胞剋隆數目降低(SH-SY5Y:24±3.07比43±6.29和IMR-32:17±2.36比37±5.41),細胞活性下降,併且細胞侵襲和遷移能力也降低(差異具有統計學意義).Western結果顯示miR-34a抑製CD44、CDK4、CCND1和CCNE2的蛋白水平.結論 miR-34a分彆通過降低CD44、CDK4、CCND1和CCNE2的錶達水平,而抑製神經母細胞瘤細胞的生長、侵襲和遷移,髮揮著腫瘤抑癌基因的作用,提示miR-34a可能作為神經母細胞瘤分子治療的靶點.
목적 검측miR-34a재원발성화전이성신경모세포류조직중적표체수평,병탐토miR-34a대신경모세포류세포생장、침습화천이적영향.방법 수집18례신경모세포류환인(신경모세포류원발부위이급전이부위),이용실시정량PCR검측miR-34a재원발성화전이성신경모세포류조직중적표체수평.재신경모세포류세포중순시전염miR-34a mimics,이용평판극륭형성실험、MTT、침습실험화체외화흔실험검측miR-34a대세포생장、침습화천이적영향.최후이용Western인적검측miR-34a대생장이급침습천이상관단백표체수평적영향.결과 여원발성신경모세포류조직상비,miR-34a재전이성신경모세포류조직중표체강저.여대조세포조상비,전염miR-34a mimics적세포극륭수목강저(SH-SY5Y:24±3.07비43±6.29화IMR-32:17±2.36비37±5.41),세포활성하강,병차세포침습화천이능력야강저(차이구유통계학의의).Western결과현시miR-34a억제CD44、CDK4、CCND1화CCNE2적단백수평.결론 miR-34a분별통과강저CD44、CDK4、CCND1화CCNE2적표체수평,이억제신경모세포류세포적생장、침습화천이,발휘착종류억암기인적작용,제시miR-34a가능작위신경모세포류분자치료적파점.
Objective To determine the expression levels of miR-34a in primary and metastatic neuroblastoma tissues and the effects of miR-34a in neuroblastoma cell growth,migration and inva sion.Methods Quantitative real-time polymerase chain reaction (Q-PCR) was performed to analyze the miR-34a expression levels in primary and metastatic neuroblastoma tissues.The cells were transfected with miR-34a mimics and controls using LipofectamineTM 2000.Then the cell growth,invasion and migration abilities were examined with colony formation,methylthiazoletetrazolium (MTT),Transwell invasion and wound healing assays.Finally the effects of miR-34a on CD44,CDK4,CCND1,CCNE2 and MYCN were measured by Western blot.Results miR-34a was down-regulated in metastatic neuroblastoma tissues.The cells with miR-34a mimics had a lower cell colony number (SHSY5Y:24 ± 3.07 vs 43 ± 6.29 & IMR-32:17-± 2.36 vs 37 ± 5.41) and reduced cell viability,invasion and migration abilities (P＜0.05).miR-34a inhibited the expressions of CD44,CDK4,CCND1 and CCNE2 proteins.Conclusions miR-34a suppresses the growth,migration and invasion of neuroblastoma cell through the inhibited expressions of CD44,CDK4,CCND1 and CCNE2.Thus it may serve as a potential molecular therapeutic target for neuroblastoma.
