馬國祥 丁茜萍 鄧海華 楊振漢

廣東深圳市寶安區(qū)福永人民醫(yī)院內(nèi)科 深圳 518103

microRNA-183通過下調(diào)Bcl-2誘導(dǎo)人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡的研究

馬國祥 丁茜萍 鄧海華 楊振漢

廣東深圳市寶安區(qū)福永人民醫(yī)院內(nèi)科 深圳 518103

目的 研究microRNA-183對神經(jīng)母細(xì)胞瘤細(xì)胞的調(diào)控作用，為神經(jīng)母細(xì)胞瘤的治療提供新策略。方法 購買microRNA-183和對照 microRNA，轉(zhuǎn)染至人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞，再使用MTT實(shí)驗(yàn)，Caspase-3活性測定試劑盒和流式檢測細(xì)胞生長和凋亡的影響。合成Bcl-2 siRNA，檢測Bcl-2對人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡的影響。轉(zhuǎn)染Bcl-2質(zhì)粒后，再轉(zhuǎn)染microRNA-183至人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞，分析Bcl-2的表達(dá)水平和人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的凋亡。結(jié)果 轉(zhuǎn)染microRNA-183降低SK-N-SH細(xì)胞的生長(P=0.005 9)，發(fā)生磷脂酰絲氨酸膜表面表達(dá)(P=0.008)和Caspase-3的激活(P=0.014)，Bcl-2的表達(dá)降低(P=0.015)。干擾SK-N-SH細(xì)胞中Bcl-2增強(qiáng)了microRNA-183誘導(dǎo)的細(xì)胞凋亡(P=0.005 8)，而過表達(dá)Bcl-2抑制了microRNA-183誘導(dǎo)的細(xì)胞凋亡(P=0.007 3)。結(jié)論 轉(zhuǎn)染microRNA-183抑制SK-N-SH細(xì)胞的生長和增殖。microRNA-183通過下調(diào)Bcl-2而誘導(dǎo)SK-N-SH細(xì)胞的凋亡，提示Bcl-2可能是神經(jīng)母細(xì)胞瘤潛在的候選治療靶點(diǎn)。

細(xì)胞凋亡；microRNA-183；Bcl-2；SK-N-SH細(xì)胞

神經(jīng)母細(xì)胞瘤嚴(yán)重威脅著患者的生活和健康[1]。神經(jīng)母細(xì)胞瘤的治療主要包括放化療、手術(shù)治療、分子靶向治療等[2]，針對不同的患者，上述治療方法療效均顯著，然而，也存在毒副作用強(qiáng)、適用人群少、分子靶點(diǎn)的選擇難等缺點(diǎn)[3-4]。microRNA調(diào)控細(xì)胞生長、凋亡和增殖作用[5]。在動物實(shí)驗(yàn)中，microRNA常被作為癌癥治療的分子靶點(diǎn)[6]。然而，microRNA-183對神經(jīng)母細(xì)胞瘤細(xì)胞生長和凋亡的調(diào)控作用尚不清楚。因此，本研究以人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞為細(xì)胞模型，探討microRNA-183對人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞可能的調(diào)控機(jī)制，以期為神經(jīng)母細(xì)胞瘤分子靶點(diǎn)的治療提供理論依據(jù)。

1 實(shí)驗(yàn)試劑和方法

1．1 實(shí)驗(yàn)試劑和實(shí)驗(yàn)細(xì)胞 小鼠源抗人Bcl-2的多克隆抗體，HRP標(biāo)記的羊源抗小鼠的IgG單克隆抗體，內(nèi)參抗actin內(nèi)參的多克隆抗體來源于美國Santa Cruz生物技術(shù)有限公司。細(xì)胞培養(yǎng)用血清和培養(yǎng)基來源于華蘭生物用品公司。MTT試劑盒來源于碧云天生物技術(shù)研究所。細(xì)胞凋亡試劑來源于北京鼎國科技有限責(zé)任公司公司。

microRNA如microRNA-83(5′-ACATCACA-TCGGTAGTTGCCAT-3′和5′- AAACAAAGAGGTAAAGGTGCA-3′)，對照microRNA(5′-CCAACGAGTGTCTTTCGAGC-3′和5′-TTGACGTTT-CACGATACAT-3′)和siRNA Bcl-2(5′-CTACGTGCTACCATGGATGC-3′和5′-TTCTAGAACTACGCTATGT-3′)，Bcl-2的質(zhì)粒來源于上海生物工程有限公司。Lipo2000轉(zhuǎn)染試劑來源于美國Invitrogen。

1．2 細(xì)胞培養(yǎng)的方法 復(fù)蘇人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞，將人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞重懸于DMEM培養(yǎng)基中，37 ℃，5% CO2[7]。

1．3 轉(zhuǎn)染方法 將microRNA-183和對照micro-RNA轉(zhuǎn)染至人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞。將人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞鋪板于24孔板，密度30%左右，將microRNA-183和對照microRNA加入lipo2000中，混勻，室溫放置7 min，將混合液加入細(xì)胞中，37 ℃，5% CO2[8-9]。

1．4 MTT試劑盒檢測細(xì)胞活性 按照MTT試劑盒的說明檢測人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的活性[10]。首先，接種人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞。按1.3的描述向人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞轉(zhuǎn)染microRNA-183或?qū)φ誱iRNA后，再加入1 mg/mL MTT反應(yīng)液，培養(yǎng)人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞24 h。最后在細(xì)胞里加入終止反應(yīng)液DMSO，終止反應(yīng)。

將6孔板人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞放入酶標(biāo)儀中，測試人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞在560 nm處的吸收值，繪制人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞生長曲線[11]。

1．5 流式細(xì)胞術(shù)檢測細(xì)胞凋亡 人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的凋亡情況使用流式檢測。首先，接種人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞。按1.3的描述向人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞轉(zhuǎn)染microRNA-183或?qū)φ誱iRNA后，向200 μL體積的細(xì)胞中加入16 μL的反應(yīng)液和2 μL的染料Annexin-V-FITC，避光反應(yīng)14 min。最后上流式檢測[12]。1．6 聚丙烯酰胺凝膠電泳和Western blot免疫印跡 首先，接種人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞。按1.3的描述向人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞轉(zhuǎn)染microRNA-183或?qū)φ誱iRNA后，收集細(xì)胞，向細(xì)胞中加入細(xì)胞裂解液裂解，制備聚丙烯酰胺凝膠電泳樣品，進(jìn)行電泳，電泳結(jié)束轉(zhuǎn)膜、封閉，孵育一抗、二抗，最后進(jìn)行顯影、定影[13]。

1．7 Caspase-3活性測試 按照試劑盒的說明檢測人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的凋亡情況。首先，接種人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞。按1.3的描述向人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞轉(zhuǎn)染microRNA-183或?qū)φ誱iRNA后，裂解細(xì)胞，向細(xì)胞樣品中加入生色底物反應(yīng)。最后在酶標(biāo)儀上測試各組樣品的光吸收值[14]，作為細(xì)胞Caspase-3活性的相對值。

2 結(jié)果

2．1 轉(zhuǎn)染microRNA-183降低了人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的活性 各實(shí)驗(yàn)組的人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞活性通過MTT測試，與轉(zhuǎn)染對照miRNA的人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞相比，轉(zhuǎn)染microRNA-183后，人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的生長被顯著抑制(P=0.005 9)。見圖1。

由于轉(zhuǎn)染對照miRNA的人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞和未轉(zhuǎn)染組人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的活性差異無統(tǒng)計(jì)學(xué)意義(P>0.05)，所以，以轉(zhuǎn)染miRNA細(xì)胞組作為對照。

圖1 轉(zhuǎn)染microRNA-183降低了人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的活性，抑制其生長 與miRNA組相比，**P<0.01 圖2 轉(zhuǎn)染microRNA-183引起人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的凋亡與miRNA組相比，**P<0.01

2．2 轉(zhuǎn)染microRNA-183引起人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的凋亡 如圖2所示，人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞磷脂酰絲氨酸的外翻，與轉(zhuǎn)染對照miRNA的人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞相比，轉(zhuǎn)染microRNA-183后，人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞膜磷脂酰絲氨酸外翻的量顯著增加(P=0.008)。

2．3 轉(zhuǎn)染microRNA-183引起人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞中Caspase-3的活化 圖3中人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞中細(xì)胞凋亡的檢測數(shù)據(jù)提示，與轉(zhuǎn)染miRNA的人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞相比，轉(zhuǎn)染microRNA-183后，人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡增加(P=0.014)。

圖3 轉(zhuǎn)染microRNA-183引起人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的Caspase-3的活化 與miRNA組相比，*P<0.05 圖4 轉(zhuǎn)染microRNA-183引起人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞中Bcl-2蛋白表達(dá)的下降

2．4 轉(zhuǎn)染microRNA-183引起人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞中Bcl-2蛋白表達(dá)的下降 如圖4所示，人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞中細(xì)胞凋亡的檢測數(shù)據(jù)提示，與轉(zhuǎn)染對照miRNA的人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞相比，轉(zhuǎn)染microRNA-183后，人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞細(xì)胞凋亡增強(qiáng)(P=0.015)。

2．5 敲低Bcl-2增強(qiáng)microRNA-183引起的人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡 如圖5所示，降低Bcl-2的表達(dá)后，再轉(zhuǎn)染microRNA-183后，人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡增加(P=0.007 6)。microRNA-183組和siBcl-2+microRNA-183細(xì)胞凋亡差異有統(tǒng)計(jì)學(xué)意義(P=0.005 8)。

圖5 敲低Bcl-2增強(qiáng)microRNA-183引起的人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡 與miRNA組相比，*P<0.05；microRNA-183組和siBcl-2+microRNA-183細(xì)胞凋亡比較，#P<0.05

2．6 轉(zhuǎn)染Bcl-2抑制了microRNA-183誘導(dǎo)的人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡 圖6中Western blot結(jié)果表明，Bcl-2的質(zhì)粒轉(zhuǎn)染增加了其水平。microRNA-183組和Bcl-2+microRNA-183組間Bcl-2比較差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。

圖6示，人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞中細(xì)胞凋亡的檢測結(jié)果表明，轉(zhuǎn)染Bcl-2增強(qiáng)Bcl-2水平后，再轉(zhuǎn)染microRNA-183后，人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡被明顯抑制(P=0.006)。microRNA-183組和Bcl-2+microRNA-183細(xì)胞凋亡差異有統(tǒng)計(jì)學(xué)意義(P=0.007 3)。

圖6 轉(zhuǎn)染Bcl-2抑制了microRNA-183誘導(dǎo)的人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡與miRNA組相比，*P<0.05；microRNA-183組和Bcl-2+microRNA-183比較，#P<0.05

3 討論

本研究以人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞為細(xì)胞模型，從蛋白水平上研究了microRNA-183對人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的調(diào)節(jié)機(jī)制，結(jié)果顯示，轉(zhuǎn)染microRNA-183降低了人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的生長，引起人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡。這與前人研究結(jié)果高度一致，即microRNA調(diào)控細(xì)胞生長和細(xì)胞凋亡[15]。

Bcl-2蛋白是Bcl-2蛋白家族中重要的細(xì)胞凋亡抑制蛋白之一[7-8]。據(jù)研究報(bào)道，Bcl-2可能受microRNA-183的調(diào)節(jié)，進(jìn)而調(diào)節(jié)人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞[9-10]。本研究結(jié)果證明，轉(zhuǎn)染microRNA-183降低了Bcl-2的蛋白水平。轉(zhuǎn)染microRNA-183和降低Bcl-2的表達(dá)后，人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的凋亡增加，而轉(zhuǎn)染Bcl-2則抑制了microRNA-183誘導(dǎo)的人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡。

本文采用不同的方法證明了Bcl-2蛋白在microRNA-183誘導(dǎo)的人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡中的作用：(1)Western blot結(jié)果顯示，microRNA-183高表達(dá)時，人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞中的Bcl-2蛋白顯著降低；(2)轉(zhuǎn)染Bcl-2 siRNA后，再轉(zhuǎn)染microRNA-183至人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞后，人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡增強(qiáng)；(3)轉(zhuǎn)染Bcl-2質(zhì)粒后，再轉(zhuǎn)染microRNA-183至人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞，人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡被顯著抑制。結(jié)果表明，Bcl-2蛋白在microRNA-183誘導(dǎo)的人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞凋亡中發(fā)揮關(guān)鍵作用，Bcl-2蛋白是潛在的神經(jīng)母細(xì)胞瘤的分子靶點(diǎn)[11-12]。Bcl-2可抑制癌細(xì)胞的細(xì)胞凋亡[13-14]，這是一致性的結(jié)論。

本文的不足和缺點(diǎn)如下：(1)缺少臨床神經(jīng)母細(xì)胞瘤的樣本和對照組織中Bcl-2蛋白水平的結(jié)果；(2)缺少預(yù)后臨床神經(jīng)母細(xì)胞瘤的樣本和對照組織中Bcl-2蛋白水平的結(jié)果；(3)缺少神經(jīng)母細(xì)胞瘤的小鼠模型和以microRNA-183為靶點(diǎn)治療的神經(jīng)母細(xì)胞瘤小鼠的療效結(jié)果。

轉(zhuǎn)染microRNA-183抑制人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的生長和增殖，microRNA-183通過下調(diào)Bcl-2而誘導(dǎo)人神經(jīng)母細(xì)胞瘤細(xì)胞株SK-N-SH細(xì)胞的凋亡，提示Bcl-2可能是神經(jīng)母細(xì)胞瘤潛在的候選治療靶點(diǎn)。

[1] Cheng M，Liu L，Lao Y，et al．MicroRNA-181a suppre-sses parkin-mediated mitophagy and sensitizes neuroblastoma cells to mitochondrial uncoupler-induced apoptosis[J]．Oncotarget，2016，7(27)：42 274-42 287．

[2] Kong B，Wu PC，Chen L，et al．microRNA-7 Protects Against 1-Methyl-4-Phenylpyridinium Iodide-Induced Cell Apoptosis in SH-SY5Y Cells by Directly Targeting Krüpple-Like Factor 4．DNA[J]．Cell Biol，2016，35(5)：217-225．

[3] Soriano A，París-Coderch L，Jubierre L，et al．Micro RNA-497 impairs the growth of chemoresistant neuroblastoma cells by targeting cell cycle，survival and vascular permeability genes[J]．Oncotarget，2016，7(8)：9 271-9 287．

[4] Nolan K，Walter F，Tuffy LP，et al．Endoplasmic reticu-lum stress-mediated upregulation of miR-29a enhances sensitivity to neuronal apoptosis[J]．Eur J Neurosci，2016，43(5)：640-652．

[5] Bevilacqua V，Gioia U，Di Carlo V，et al．Identification of linc-NeD125，a novel long non coding RNA that hosts miR-125b-1 and negatively controls proliferation of human neuroblastoma cells[J]．RNA Biol，2015， 12(12)：1 323-1 337．

[6] Bachetti T，Di Zanni E，Ravazzolo R，et al．miR-204 mediates post-transcriptional down-regulation of PHOX2B gene expression in neuroblastoma cells[J]．Biochim Biophys Acta，2015，1 849(8)：1 057-1 065．

[7] Wu K，Yang L，Chen J，et al．miR-362-5p inhibits proliferation and migration of neuroblastoma cells by targeting phosphatidylinositol 3-kinase-C2β[J]．FEBS Lett，2015，589(15)：1 911-1 919．

[8] Carpenter D，Hsiang C，Jiang X，et al．The herpes simplex virus type 1 (HSV-1)latency-associated transcript (LAT)protects cells against cold-shock-induced apoptosis by maintaining phosphorylation of protein kinase B (AKT)[J]．J Neurovirol，2015，21(5)：568-575．

[9] Zhang W，Zhao L，Liu J，et al．Cisplatin induces platelet apoptosis through the ERK signaling pathway[J]．Thromb Res，2012，130(1)：81-91．

[10] Tao Z，Zhao H，Wang R，et al．Neuroprotective effect of microRNA-99a against focal cerebral ischemia-reperfusion injury in mice[J]．J Neurol Sci，2015，355(1/2)：113-119．

[11] Du X，Wang H，Xu F，et al．Enterovirus 71 induces apoptosis of SH．SY5Y human neuroblastoma cells through stimulation of endogenous microRNA let-7b expression[J]．Mol Med Rep，2015，12(1)：953-959．

[12] Rihani A，Van Goethem A，Ongenaert M，et al．Genome wide expression profiling of p53 regulated miRNAs in neuroblastoma[J]．Sci Rep，2015，5：9 027．

[13] Zhao Z，Ma X，Sung D，et al．microRNA-449a functions as a tumor suppressor in neuroblastoma through inducing cell differentiation and cell cycle arrest[J]．RNA Biol，2015，12(5)：538-554．

[14] Ren X，Bai X，Zhang X，et al．Quantitative nuclear proteomics identifies that miR-137-mediated EZH2 reduction regulates resveratrol-induced apoptosis of neuroblastoma cells[J]．Mol Cell Proteomics，2015，14(2)：316-328．

[15] Harvey H，Piskareva O，Creevey L，et al．Modulation of chemotherapeutic drug resistance in neuroblastoma SK-N-AS cells by the neural apoptosis inhibitory protein and miR-520f[J]．Int J Cancer，2015，136(7)：1 579-1 588．

(收稿2016-10-20)

microRNA-183 induces apoptosis of human neuroblastoma cell line SK-N-SH by down regulating Bcl-2 disease rats

MaGuoxiang，DingQianping，DengHaihua，YangZhenhan

FuyongPeople'sHospitalofShenzhenBaoanDistrict，Shenzhen518103，China

Objective To study the regulation of microRNA-183 on neuroblastoma cells and to provide a new strategy for the treatment of neuroblastoma．Methods microRNA and microRNA-183 were transfected into SK-N-SH cells，and then MTT experiment and Caspase-3 activity assay and flow cytometry were employed to investigate the role of microRNA-183 in growth and apoptosis of human neuroblastoma cell line SK-N-SH．Bcl-2 siRNA was transfected into SK-N-SH cells，and effects of Bcl-2 on human neuroblastoma cell line SK-N-SH cells apoptosis was tested．After transfection of Bcl-2 plasmid，microRNA-183 was transfected into human neuroblastoma cell line SK-N-SH cells，and the expression level of Bcl-2 and apoptosis of human neuroblastoma cell line SK-N-SH were analyzed．Results The transfection of microRNA-183 reduced cell growth of the SK-N-SH cells (P=0.005 9)，and the expression of phosphatidylserine on the membrane (P=0.008) and activation of Caspase-3 were observed (P=0.014)，and the expression of Bcl-2 was decreased (P=0.015)．Bcl-2 interference of human neuroblastoma cell lines enhances apoptosis of the human neuroblastoma cell line SK-N-SH induced by microRNA-183 (P=0.005 8)，and overexpression of Bcl-2 inhibits apoptosis induced by microRNA-183 (P=0.007 3)．Conclusion The transfection of microRNA-183 inhibits the growth and proliferation of human neuroblastoma cell line SK-N-SH．MicroRNA-183 induces apoptosis of SK-N-SH by down regulating Bcl-2，which suggests that Bcl-2 may be a potential candidate therapeutic target for neuroblastoma．

Apoptosis；microRNA-183；Bcl-2；SK-N-SH cell

R730.264

A

1673-5110(2017)07-0003-04