中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2015年
4期
326-330
,共5页
Raji细胞%抗药性%基因,PHB%微RNAs
Raji細胞%抗藥性%基因,PHB%微RNAs
Raji세포%항약성%기인,PHB%미RNAs
Raji cells%Drug resistance%Gene,PHB%MicroRNAs
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Raji/A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一般形态并计数,采用RT-PCR方法检测Raji/A及Raji/S细胞中PHB mRNA与miR-27a的表达水平,采用Western blot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48 h,Raji/A及Raji/S细胞的半数抑制浓度(IC50)分别为344.17及1.12 μg/ml,耐药倍数为306.47倍.Raji/A与Raji/S细胞经4μg/ml ADR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).PHBl mRNA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);PHB2 mRNA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).PHBl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.
目的 建立阿黴素(ADR)耐藥Raji細胞株(Raji/A),檢測線粒體抗增殖蛋白(Prohibitin,PHB)及微小RNA-27a(miR-27a)在Raji細胞及其敏感株(Raji/S)中的錶達變化併探討其臨床意義.方法 建立Raji/A細胞株,採用CCK-8法檢測細胞增殖抑製率,採用相差顯微鏡觀察細胞一般形態併計數,採用RT-PCR方法檢測Raji/A及Raji/S細胞中PHB mRNA與miR-27a的錶達水平,採用Western blot法檢測PHB1蛋白在Raji/A及Raji/S細胞線粒體中的錶達水平.結果 不同濃度ADR處理48 h,Raji/A及Raji/S細胞的半數抑製濃度(IC50)分彆為344.17及1.12 μg/ml,耐藥倍數為306.47倍.Raji/A與Raji/S細胞經4μg/ml ADR處理24h,凋亡率分彆為(13.48±2.20)％及(69.89±1.94)％,差異有統計學意義(P＜0.05).PHBl mRNA在Raji/A細胞中的錶達明顯高于Raji/S細胞,為(7.66±3.27)倍(P＜0.05);PHB2 mRNA在Raji/A細胞中的錶達是Raji/S的(1.25±0.47)倍,差異無統計學意義(P＞0.05);miR-27a在Raji/A細胞中的錶達明顯高于Raji/S細胞,為(128.40±31.59)倍(P＜0.05).PHBl蛋白在Raji/A及Raji/S細胞線粒體中的相對錶達量分彆為9.87±0.22及1.96±0.06(P＜0.05).結論 成功構建穩定Raji/A細胞模型,PHB1及miR-27a在Raji/A細胞中錶達升高,其高錶達可能參與Raji細胞ADR耐藥機製.
목적 건립아매소(ADR)내약Raji세포주(Raji/A),검측선립체항증식단백(Prohibitin,PHB)급미소RNA-27a(miR-27a)재Raji세포급기민감주(Raji/S)중적표체변화병탐토기림상의의.방법 건립Raji/A세포주,채용CCK-8법검측세포증식억제솔,채용상차현미경관찰세포일반형태병계수,채용RT-PCR방법검측Raji/A급Raji/S세포중PHB mRNA여miR-27a적표체수평,채용Western blot법검측PHB1단백재Raji/A급Raji/S세포선립체중적표체수평.결과 불동농도ADR처리48 h,Raji/A급Raji/S세포적반수억제농도(IC50)분별위344.17급1.12 μg/ml,내약배수위306.47배.Raji/A여Raji/S세포경4μg/ml ADR처리24h,조망솔분별위(13.48±2.20)％급(69.89±1.94)％,차이유통계학의의(P＜0.05).PHBl mRNA재Raji/A세포중적표체명현고우Raji/S세포,위(7.66±3.27)배(P＜0.05);PHB2 mRNA재Raji/A세포중적표체시Raji/S적(1.25±0.47)배,차이무통계학의의(P＞0.05);miR-27a재Raji/A세포중적표체명현고우Raji/S세포,위(128.40±31.59)배(P＜0.05).PHBl단백재Raji/A급Raji/S세포선립체중적상대표체량분별위9.87±0.22급1.96±0.06(P＜0.05).결론 성공구건은정Raji/A세포모형,PHB1급miR-27a재Raji/A세포중표체승고,기고표체가능삼여Raji세포ADR내약궤제.
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 of phb mRNA and miR-27a in ADR-resistant cells (Raji/A) and sensitive cells (Raji/S) via realtime quantitative polymerase chain reaction (RT-PCR).Results The ADR-resistant Raji cell lines were built;expression of PHB 1 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).Conclusion By building the experimental model of Raji ADR-resistant cell lines,high expression level of PHB1 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.
