摘要
目的 观察MBD1通过对mdr1转录区域结合位点甲基化影响,在转录水平间接调控胰腺癌mdr1基因表达.方法 应用RNA干扰技术对人胰腺癌细胞株BxPC-3中MBD1基因进行抑制;应用定量荧光聚合酶链式反应（FQ-PCR）技术、MSP（甲基化专用聚合酶链式反应）方法分别检测转染前后细胞株中的mdr1 mRNA的表达差异和转录区域结合位点甲基化变化.结果 成功构建并转染MBDI siRNAs至人原位胰腺腺癌细胞BxPC-3（BxPC-3）细胞中,证实MBDI mRNA水平明显下调（下调幅度为93.19%）.MBD1下调后,mdr1 DNA转录区域（-110GC和-50GC）甲基化分别上调（上调幅度分别为181.98%和409.57%）;mdr1 mRNA表达明显下调（下调幅度为97.79%）.结论 MBD1可通过对mdr1转录区域结合位点甲基化的影响,在转录水平间接调控胰腺癌mdr1基因表达.
Objective To investigate that methyl-CpG binding domain protein 1 （MBD1） could regulate multidrug resistance gene 1 （mdr1） during its RNA transcription indirectly by influencing the methylation of mdr1 transcription domain. Methods MBD1 gene in human pancreatic cancer cell line BxPC-3 was depressed by RNA interference （RNAi）. The expression of mdr1 mRNA and the methylation of its transcription domain were detected by the techniques of real-time fluorescent quantitative polymerase chain reaction （FQ-PCR） and methylations expression and metspecific PCR （MSP） respectively before and after transfection. Results MBD1 siRNAs were constructed and transfected into BxPC-3 successfully. The expression of MBD1 mRNA was down-regulated obviously （93. 19% ）. The methylation of-1 10GC-box and-50GC-box was up-regulated （181. 98% and 409. 57% respectively） and the expression of mdr1 mRNA was down-regulated （97.79%） accordingly. Conclusion MBD1 could regulate the expression of mdr1 indirectly by influencing the methylation of mdr1 transcription domain during its RNA transcription.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2010年第6期763-765,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目（30672057）.
关键词
胰腺癌
多药耐药基因
转录
甲基化
Pancreatic carcinoma
Multidrug resistant gene
Transcription
Methylation
