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
333-337
,共5页
杨少波%郑继翠%郑珊%董岿然%肖现民
楊少波%鄭繼翠%鄭珊%董巋然%肖現民
양소파%정계취%정산%동규연%초현민
神经母细胞瘤%性别决定区Y蛋白质%细胞增殖%细胞分化
神經母細胞瘤%性彆決定區Y蛋白質%細胞增殖%細胞分化
신경모세포류%성별결정구Y단백질%세포증식%세포분화
Neuroblastoma%Sex-determining region Y protein%Cell proliferation%Cell differentiation
目的 探讨胚胎干细胞因子Sox2对神经母细胞瘤Ⅰ型细胞增殖分化的影响.方法 应用慢病毒载体介导RNA干扰处理Ⅰ型细胞BE(2)-C构建Sox2低表达细胞株,采用CCK-8活细胞数检测,分析Sox2对BE(2)-C细胞增殖能力的影响.维甲酸和5-溴2'-去氧尿甙干预BE(2)-C细胞诱导分化后采用Western blot检测细胞标志蛋白的表达差异,分析Sox2对BE(2)-C细胞分化能力的影响.结果 CCK-8活细胞数检测显示72 h时Sox2 shRNA组的细胞AV值(0.72±0.18)低于空白对照组(1.24±0.21)和阴性对照组(1.16±0.26),差异有统计学意义(P＜0.05).RA或BrdU诱导分化后Western blot检测细胞标志蛋白(Peripherin、NF-68、Vimentin、S-100)显示,经RA或BrdU诱导分化的Sox2 shRNA组细胞其标志蛋白表达量均高于经同种药物诱导的空白对照组(BE(2)-C)和阴性对照组(Sox2 shRNA空载体)细胞,差异有统计学意义(P＜0.05).结论 Sox2促进神经母细胞瘤Ⅰ型细胞的增殖,但抑制其向低度恶性细胞分化,提示Sox2有望成为神经母细胞瘤基因治疗的新靶点.
目的 探討胚胎榦細胞因子Sox2對神經母細胞瘤Ⅰ型細胞增殖分化的影響.方法 應用慢病毒載體介導RNA榦擾處理Ⅰ型細胞BE(2)-C構建Sox2低錶達細胞株,採用CCK-8活細胞數檢測,分析Sox2對BE(2)-C細胞增殖能力的影響.維甲痠和5-溴2'-去氧尿甙榦預BE(2)-C細胞誘導分化後採用Western blot檢測細胞標誌蛋白的錶達差異,分析Sox2對BE(2)-C細胞分化能力的影響.結果 CCK-8活細胞數檢測顯示72 h時Sox2 shRNA組的細胞AV值(0.72±0.18)低于空白對照組(1.24±0.21)和陰性對照組(1.16±0.26),差異有統計學意義(P＜0.05).RA或BrdU誘導分化後Western blot檢測細胞標誌蛋白(Peripherin、NF-68、Vimentin、S-100)顯示,經RA或BrdU誘導分化的Sox2 shRNA組細胞其標誌蛋白錶達量均高于經同種藥物誘導的空白對照組(BE(2)-C)和陰性對照組(Sox2 shRNA空載體)細胞,差異有統計學意義(P＜0.05).結論 Sox2促進神經母細胞瘤Ⅰ型細胞的增殖,但抑製其嚮低度噁性細胞分化,提示Sox2有望成為神經母細胞瘤基因治療的新靶點.
목적 탐토배태간세포인자Sox2대신경모세포류Ⅰ형세포증식분화적영향.방법 응용만병독재체개도RNA간우처리Ⅰ형세포BE(2)-C구건Sox2저표체세포주,채용CCK-8활세포수검측,분석Sox2대BE(2)-C세포증식능력적영향.유갑산화5-추2'-거양뇨대간예BE(2)-C세포유도분화후채용Western blot검측세포표지단백적표체차이,분석Sox2대BE(2)-C세포분화능력적영향.결과 CCK-8활세포수검측현시72 h시Sox2 shRNA조적세포AV치(0.72±0.18)저우공백대조조(1.24±0.21)화음성대조조(1.16±0.26),차이유통계학의의(P＜0.05).RA혹BrdU유도분화후Western blot검측세포표지단백(Peripherin、NF-68、Vimentin、S-100)현시,경RA혹BrdU유도분화적Sox2 shRNA조세포기표지단백표체량균고우경동충약물유도적공백대조조(BE(2)-C)화음성대조조(Sox2 shRNA공재체)세포,차이유통계학의의(P＜0.05).결론 Sox2촉진신경모세포류Ⅰ형세포적증식,단억제기향저도악성세포분화,제시Sox2유망성위신경모세포류기인치료적신파점.
Objective To explore the effects of Sox2 on cell proliferation and differentiation of Ⅰtype neuroblastoma cell.Methods Sox2 mRNA knockdown BE (2)-C cell line was established by RNA interference via lentiviral vectors.Cell counting kit-8 (CCK-8) assay was applied to test the effect of Sox2 on the proliferation of BE(2)-C cell.Western blot was performed to detect the marker proteins after induced differentiation in order to analyze the effect of Sox2 on differentiation properties.Results The absorbance value (AV) of Sox2 shRNA cells(0.72 ± 0.18)was lower than BE(2)-C cells (1.24 ± 0.21)and shRNA control cells(1.16 ± 0.26)at 72 h in CCK-8 assay (P＜0.05).Sox2 shRNA cells exhibited the elevated expression levels of marker proteins (Peripherin,NF-68,Vimentin & S100)of N or S-type cell than BE(2)-C and shRNA control cells after induced differentiation (P＜0.05).Conclusions Sox2 promotes cell proliferation and inhibits the differentiation of Ⅰ-type neuroblastoma cell.It may serve as a new target of gene therapy for neuroblastoma.
