白血病·淋巴瘤
白血病·淋巴瘤
백혈병·림파류
JOURNAL OF LEUKEMIA & LYMPHOMA
2014年
3期
148-151,155
,共5页
孝飞飞%贾秀红%李建厂%李有杰
孝飛飛%賈秀紅%李建廠%李有傑
효비비%가수홍%리건엄%리유걸
肿瘤%Apollon基因%RNA,小分子干扰%MCF-7细胞
腫瘤%Apollon基因%RNA,小分子榦擾%MCF-7細胞
종류%Apollon기인%RNA,소분자간우%MCF-7세포
Neoplasms%Apollon gene%RNA,small interfering%MCF-7 cell
目的 筛选干扰Apollon基因表达的siRNA序列并鉴定其功能.方法 采用化学方法合成针对Apollon基因不同位点的siRNA序列;应用Lipofectamine 2000转染人类乳腺癌MCF-7细胞;反转录PCR(RT-PCR)检测Apollon mRNA表达水平;细胞免疫荧光结合激光共聚焦技术定量分析Apollon蛋白的表达;四甲基偶氮唑蓝(MTT)和流式细胞术分别检测siRNA干扰Apollon后对MCF-7细胞增殖和凋亡的影响.结果 合成的3对siRNA序列均能抑制Apollon mRNA表达,其siRNA1、siRNA2、siRNA3对ApollonmRNA的抑制率分别为(36.201±11.629)％、(67.308±7.686)％、(47.123±12.000)％,与对照组相比差异均有统计学意义(均P＜ 0.05);siRNA2转染MCF-7细胞后Apollon蛋白的荧光强度为(14.97±2.08)％,增殖抑制率达(73.361±2.118)％,凋亡率为(28.793±0.743)％.结论 筛选出的siRNA2序列可有效沉默Apollon基因的表达,显著抑制MCF-7细胞的增殖和促进其凋亡,为肿瘤靶向治疗提供实验依据.
目的 篩選榦擾Apollon基因錶達的siRNA序列併鑒定其功能.方法 採用化學方法閤成針對Apollon基因不同位點的siRNA序列;應用Lipofectamine 2000轉染人類乳腺癌MCF-7細胞;反轉錄PCR(RT-PCR)檢測Apollon mRNA錶達水平;細胞免疫熒光結閤激光共聚焦技術定量分析Apollon蛋白的錶達;四甲基偶氮唑藍(MTT)和流式細胞術分彆檢測siRNA榦擾Apollon後對MCF-7細胞增殖和凋亡的影響.結果 閤成的3對siRNA序列均能抑製Apollon mRNA錶達,其siRNA1、siRNA2、siRNA3對ApollonmRNA的抑製率分彆為(36.201±11.629)％、(67.308±7.686)％、(47.123±12.000)％,與對照組相比差異均有統計學意義(均P＜ 0.05);siRNA2轉染MCF-7細胞後Apollon蛋白的熒光彊度為(14.97±2.08)％,增殖抑製率達(73.361±2.118)％,凋亡率為(28.793±0.743)％.結論 篩選齣的siRNA2序列可有效沉默Apollon基因的錶達,顯著抑製MCF-7細胞的增殖和促進其凋亡,為腫瘤靶嚮治療提供實驗依據.
목적 사선간우Apollon기인표체적siRNA서렬병감정기공능.방법 채용화학방법합성침대Apollon기인불동위점적siRNA서렬;응용Lipofectamine 2000전염인류유선암MCF-7세포;반전록PCR(RT-PCR)검측Apollon mRNA표체수평;세포면역형광결합격광공취초기술정량분석Apollon단백적표체;사갑기우담서람(MTT)화류식세포술분별검측siRNA간우Apollon후대MCF-7세포증식화조망적영향.결과 합성적3대siRNA서렬균능억제Apollon mRNA표체,기siRNA1、siRNA2、siRNA3대ApollonmRNA적억제솔분별위(36.201±11.629)％、(67.308±7.686)％、(47.123±12.000)％,여대조조상비차이균유통계학의의(균P＜ 0.05);siRNA2전염MCF-7세포후Apollon단백적형광강도위(14.97±2.08)％,증식억제솔체(73.361±2.118)％,조망솔위(28.793±0.743)％.결론 사선출적siRNA2서렬가유효침묵Apollon기인적표체,현저억제MCF-7세포적증식화촉진기조망,위종류파향치료제공실험의거.
Objective To screen siRNAs that can effectively inhibit Apollon gene expression and determine the cellular functions of those siRNAs.Methods A chemical synthesis method was used to synthesize 3 siRNA sequences against different sites of Apollon.They were transfected into the human breast cancer MCF-7 cells by using Lipofectamine 2000.mRNA level of Apollon was determined by reverse transcription-polymerase chain reaction (RT-PCR).Cellular immunity fluorescence quantitative analysis combined with confocal laser technology was used to determine the protein level of Apollon.Methyl thiazolyl tetrazolium bromide (MTT) assay and flow cytometry were used to determine the effects of siRNA targeting Apollon on proliferation and apoptosis of MCF-7 cells,respectively.Results Three pairs of siRNA could significantly inhibit Apollon mRNA expression,at the inhibition rates of (36.201±11.629) ％,(67.308±7.686) ％and (47.123±12.000) ％,respectively (P ＜ 0.05).After tranfection by siRNA2,Apollon protein fluorescence intensity was (14.97±2.08) ％ compared with control cells.The cell proliferation MCF-7 was inhibited by (73.361±2.118) ％and apoptosis was increased by (28.793±0.743) ％.Conclusions Screened siRNA2 effectively silences Apollon gene expression,effectively inhibits the proliferation and increases the apoptosis of MCF-7 cells.This provids the foundation for its clinical application in cancer therapy.
