SDF-1α和c-MYC蛋白调控速激肽受体-1截短型mRNA表达的研究
The study on SDF-1α and c-MYC protein regulate tachykinin receptor 1-trucated mRNA expression
摘要目的 观察基质细胞衍生因子1α(stromal cell derived factor 1α,SDF-1α)、c-MYC蛋白对速激肽受体-1截短型(tachykinin receptor 1 trucated,TACRI-Tr)转录水平的影响.方法 构建c-myc基因shRNA表达载体;用小干扰RNA方法沉默乳腺癌细胞MCF-7的c-myc基因,分成c-myc+细胞组和c-myc-细胞组,相关实验组加SDF-1α细胞因子中和抗体,用实时定量PCR检测TYACR1-Tr mRNA表达水平.结果 c-myc基因shRNA表达载体构建成功;在正常培养条件下,c-myc-细胞组的TACR1-Tr转录水平低于c-myc+细胞组(P<0.05);加了SDF-1α中和抗体后,c-myc-组TACR1-Tr的mRNA表达水平高于c-myc+组(P<0.05).结论 MCF-7细胞在正常培养条件下,c-MYC蛋白可以反式激活TACRI-Tr的转录;但加入SDF-1α因子中和抗体后,c-MYC蛋白失去了这种激活作用.
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abstractsObjective To investigate how SDF-1α and c-MYC protein regulates TACRl-Tr expression. Methods c-myc shRNA vector was constructed, small interfering RNA was employed for silencing c-myc gene in MCF-7 breast cancer cell. SDF-1α neutralized antibody was used in c-myc+ cell group and c-myc- cell group, while other c-myc+ cell group and c-myc- cells group were cultured under normal condition. The mRNA level of TACRl-Tr was determined by real-time PCR. Results c-myc shRNA vector was constructed successfully, in the normal presence of SDF-la, the level of TACRl-Tr mRNA in c-myc- cell group were lower than that in c-myc+ cell group( P < 0.05). But in the presence of SDF-la neutralized antibody, TACRl-Tr mRNA level of c-myc- cell group was higher than that of c-myc+ cell group(P < 0.05). Conclusion In the normal culture condition, c-MYC protein may transactivate TACRl-Tr transcription in MCF-7 cell, in the presence of SDF-1α neutral antibody, c-MYC protein lost the activity of transactivating for TACRl-Tr transciption.
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