CAJ | 학술논문

目的探讨microRNA-92 (miR-92)对神经母细胞瘤细胞增殖的促进作用,并预测miR-92的靶基因.方法人工设计合成miR-92的反义寡核苷酸(anti-miR-92寡核苷酸),利用脂质体LipofectamineTM 2000转染神经母细胞瘤SK-N-SH和SK-N-BE2细胞系,利用实时定量PCR检测转染后细胞内miR-92的表达水平,并通过噻唑蓝(MTT)比色法和流式细胞仪检测细胞的增殖和凋亡情况;利用生物信息学方法对miR-92的候选靶基因进行预测.结果与对照组细胞相比,两种神经母细胞瘤细胞转染anti-miR-92后,miR-92的水平降低约70％,MTT检测细胞的活性降低约20％,流式分析仪检测发现增殖的细胞数降低约50％～60％,而细胞凋亡数增加约2～3倍,差异均具有统计学意义(P＜0.05).通过生物信息学和对靶基因潜在功能分析,初步选定BCL2-like 11(BCL2L11)和phosphatase and tensin homolog deleted on chromosome ten (PTEN)作为是miR-92的候选靶基因.结论抑制miR-92的表达导致神经母细胞瘤细胞的增殖降低,凋亡增加,从而可能进一步影响神经母细胞瘤的生长,miR-92可能具有作为临床上神经母细胞瘤治疗分子标记物的潜在功能,为以后更有意义的分子治疗奠定基础.
목적 탐토microRNA-92 (miR-92)대신경모세포류세포증식적촉진작용,병예측miR-92적파기인.방법 인공설계합성miR-92적반의과핵감산(anti-miR-92과핵감산),이용지질체LipofectamineTM 2000전염신경모세포류SK-N-SH화SK-N-BE2세포계,이용실시정량PCR검측전염후세포내miR-92적표체수평,병통과새서람(MTT)비색법화류식세포의검측세포적증식화조망정황;이용생물신식학방법대miR-92적후선파기인진행예측.결과 여대조조세포상비,량충신경모세포류세포전염anti-miR-92후,miR-92적수평강저약70％,MTT검측세포적활성강저약20％,류식분석의검측발현증식적세포수강저약50％～60％,이세포조망수증가약2～3배,차이균구유통계학의의(P＜0.05).통과생물신식학화대파기인잠재공능분석,초보선정BCL2-like 11(BCL2L11)화phosphatase and tensin homolog deleted on chromosome ten (PTEN)작위시miR-92적후선파기인.결론 억제miR-92적표체도치신경모세포류세포적증식강저,조망증가,종이가능진일보영향신경모세포류적생장,miR-92가능구유작위림상상신경모세포류치료분자표기물적잠재공능,위이후경유의의적분자치료전정기출.
Objective To investigate the effects of miR-92 on the proliferation and apoptosis in human neuroblastoma (NB) cells,and to predict its putative targets.Methods Antisense oligonucleotides of miR-92 were synthesized and transfected into SK-N-SH and SK-N-BE2 cells,then real time PCR was performed for the detection of miR-92 expression.Cell proliferation and apoptosis were determined using MTT assay and flow cytometry.The putative targets for miR-92 were predicted using bioinformatics.Results Cell viability and proliferation of anti-miR-92 transfected cells were reduced by 20％ and 50％-60％,respectively,while apoptosis increased by 2-3 fold (P＜0.05).Using bioinformatics,BCL2-like 11 (apoptosis facilitator) (BCL2L11) and PTEN were selected as the candidate targets for miR-92.Conclusions Our data indicate that the inhibition of miR-92 can suppress NB cell proliferation and increase cell apoptosis,suggesting miR-92 may play important roles in NB growth.In addition,miR-92 may serve as a novel biomarker for therapeutic strategy of NB patients.