阿霉素耐药Raji细胞株的建立与Prohibitin及微小RNA-27a的表达情况分析  

The establishment of Raji drug-resistant cell line and analyses of prohibitin and miR-27a expression

作　　者：郑荣立[1] 姜玉杰[1] 王欣[1] 

机构地区：[1]山东大学附属省立医院血液科,济南250021

出　　处：《中华血液学杂志》2015年第4期326-330,共5页Chinese Journal of Hematology

基　　金：基金项目：国家自然科学基金（81270598、31340009）;山东省自然科学基金（ZR2012HZ003、ZR2011HQ009）

摘　　要：目的建立阿霉素（ADR）耐药Raji细胞株（Raji／A），检测线粒体抗增殖蛋白（Prohibitin，PHB）及微小RNA-27a（miR-27a）在Raji细胞及其敏感株（Raji／S）中的表达变化并探讨其临床意义。方法建立RaNA细胞株，采用CCK．8法检测细胞增殖抑制率，采用相差显微镜观察细胞-般形态并计数，采用RT-PCR方法检测Raji／A及Raji／S细胞中PHBmRNA与miR．27a的表达水平，采用Westernblot法检测PHB1蛋白在Raji／A及Raji／S细胞线粒体中的表达水平。结果不同浓度ADR处理48h，Raji／A及R@／S细胞的半数抑制浓度（IC50）分别为344．17及1．12gg／ml，耐药倍数为306．47倍。Raji／A与Raji／S细胞经4pg／mlADR处理24h，凋亡率分别为（13．48±2．20）％及（69．89±1．94）％，差异有统计学意义（P〈0．05）。PHB1mRNA在Raji／A细胞中的表达明显高于Raji／S细胞，为（7．66±3．27）倍（P〈0．05）；PHB2mRNA在Raji／A细胞中的表达是Raji／S的（1．25±0．47）倍，差异无统计学意义（P〉0．05）；miR-27a在Raji／A细胞中的表达明显高于Raji／S细胞，为（128．40±31．59）倍（P〈0．05）。PHB1蛋白在Raji／A及Raji／S细胞线粒体中的相对表达量分别为9．87±0．22及1．96±0．06（P〈0．05）。结论成功构建稳定Raji／A细胞模型，PHB1及miR-27a在Raji／A细胞中表达升高，其高表达可能参与Raji细胞ADR耐药机制。Objective To establish Raji adriamycin （ADR）-resistant cell lines and analysis the expression of mitochondria Prohibitin （PHB） and microRNA-27a （miR-27a）, as well as discuss its clinical significance. Methods Built ADR-resistant Raji cells, detected their resistant index and drug-resistant spectrum and stability, observed their morphology and growth characteristics in general; evaluated the expression ofphb mRNA and miR-27a in ADR-resistant cells （Raji/A） and sensitive cells （Raji/S） via real- time quantitative polymerase chain reaction （RT-PCR）. Results The ADR-resistant Raji cell lines were built; expression of PHB1 mRNA in Raji/A was evidently higher than in Raji/S （P〈0.05）, the expressing difference of PHB2 mRNA in Raji/A and Raji/S was statistically meaningless （P〉0.05）, the expression of miR-27a in Raji/A was much higher than in Raji/S （P〈0.05）. Conelusion By building the experimental model of Raji ADR-resistant cell lines, high expression level of PHBI and miR-27a were detected in the cell lines, indicating that PHB 1 and miR-27a may be associated with ADR-resistance of Raji cells.

关 键 词：RAJI细胞 抗药性 基因 PHB 微RNAS 

分 类 号：R96[医药卫生—药理学]