中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2010年
6期
361-365
,共5页
白血病%抗药性,多药%微RNAs%K562/A02细胞
白血病%抗藥性,多藥%微RNAs%K562/A02細胞
백혈병%항약성,다약%미RNAs%K562/A02세포
Leukemia%Drug resistance,multiple%MicroRNA%K562/A02 cells
目的 通过检测慢性粒细胞白血病急变细胞系K562及其阿霉素耐药株K562/A02的微小RNA(microRNA、miR)表达差异,探讨microRNA与白血病化疗耐药的关系.方法 MTT法检测K562/A02及其亲本细胞系K562的耐药性能;流式细胞术检测K562与K562/A02细胞的P-gp表达;运用microRNA芯片技术筛查K562与K562/A02细胞之间差异表达的microRNA,随后用实时荧光定量RT-PCR方法进一步证实.结果 阿霉素耐药株K562/A02相对于其亲本细胞系K562对阿霉素的耐药倍数为180倍;K562细胞P-gp的表达率为0.2%,K562/A02细胞P-gp的表达率为86%;microRNA芯片结果显示K562/A02与K562细胞之间有22种microRNA表达存在显著的差异(P<0.01),表达差异在2倍以上的有9种,其中miR-221、miR-155、miR-451在K562/A02细胞表达上调,而miR-98、miR-181a、let-7f、miR-424、let-7g和miR-563则表达下调.实时荧光定量RT-PCR进一步证实了上述结果,并显示miR-451、miR-155、miR-221、let-7f、miR-424在两种细胞中表达差异显著.结论 K562/A02与K562细胞存在microRNA表达差异,其中miR-451、miR-155和miR-221在K562/A02中表达显著上调,而let-7f、miR-424则显著下调,提示microRNA可能参与白血病耐药形成,差异表达的microRNA可能为逆转白血病耐药提供新的作用靶点.
目的 通過檢測慢性粒細胞白血病急變細胞繫K562及其阿黴素耐藥株K562/A02的微小RNA(microRNA、miR)錶達差異,探討microRNA與白血病化療耐藥的關繫.方法 MTT法檢測K562/A02及其親本細胞繫K562的耐藥性能;流式細胞術檢測K562與K562/A02細胞的P-gp錶達;運用microRNA芯片技術篩查K562與K562/A02細胞之間差異錶達的microRNA,隨後用實時熒光定量RT-PCR方法進一步證實.結果 阿黴素耐藥株K562/A02相對于其親本細胞繫K562對阿黴素的耐藥倍數為180倍;K562細胞P-gp的錶達率為0.2%,K562/A02細胞P-gp的錶達率為86%;microRNA芯片結果顯示K562/A02與K562細胞之間有22種microRNA錶達存在顯著的差異(P<0.01),錶達差異在2倍以上的有9種,其中miR-221、miR-155、miR-451在K562/A02細胞錶達上調,而miR-98、miR-181a、let-7f、miR-424、let-7g和miR-563則錶達下調.實時熒光定量RT-PCR進一步證實瞭上述結果,併顯示miR-451、miR-155、miR-221、let-7f、miR-424在兩種細胞中錶達差異顯著.結論 K562/A02與K562細胞存在microRNA錶達差異,其中miR-451、miR-155和miR-221在K562/A02中錶達顯著上調,而let-7f、miR-424則顯著下調,提示microRNA可能參與白血病耐藥形成,差異錶達的microRNA可能為逆轉白血病耐藥提供新的作用靶點.
목적 통과검측만성립세포백혈병급변세포계K562급기아매소내약주K562/A02적미소RNA(microRNA、miR)표체차이,탐토microRNA여백혈병화료내약적관계.방법 MTT법검측K562/A02급기친본세포계K562적내약성능;류식세포술검측K562여K562/A02세포적P-gp표체;운용microRNA심편기술사사K562여K562/A02세포지간차이표체적microRNA,수후용실시형광정량RT-PCR방법진일보증실.결과 아매소내약주K562/A02상대우기친본세포계K562대아매소적내약배수위180배;K562세포P-gp적표체솔위0.2%,K562/A02세포P-gp적표체솔위86%;microRNA심편결과현시K562/A02여K562세포지간유22충microRNA표체존재현저적차이(P<0.01),표체차이재2배이상적유9충,기중miR-221、miR-155、miR-451재K562/A02세포표체상조,이miR-98、miR-181a、let-7f、miR-424、let-7g화miR-563칙표체하조.실시형광정량RT-PCR진일보증실료상술결과,병현시miR-451、miR-155、miR-221、let-7f、miR-424재량충세포중표체차이현저.결론 K562/A02여K562세포존재microRNA표체차이,기중miR-451、miR-155화miR-221재K562/A02중표체현저상조,이let-7f、miR-424칙현저하조,제시microRNA가능삼여백혈병내약형성,차이표체적microRNA가능위역전백혈병내약제공신적작용파점.
Objective To explore the relationship between micmRNA and drug resistance in leukemia treatment by screening and identifying the microRNAs which ditierentially express in K562 cell line and its adriamycin resistant cells-K562/A02 cell line.Methods The drug resistance potency of K562/A02 cells was evaluated by M,TT assay.P-gp expression of K562 and K562/A02 cells were detected by flow cytometry (FCM).The differentially expressed microRNAs in K562 and K562/A02 cells were analyzed by microarray technique and Real Time RT-PCR.Results The resistance to adriamycin (ADM) of K562/A02 cells was 180 fold greater than that of K562 cells.P-gp expression rate of K562 and K562/A02 cells was 0.2% and 86%,respectively.Twenty-two microRNAs expressed difierentially in K562 and K562/A02 cells(P<0.01).As compared to K562 cells,expressions of miR-221,miR-155 and miR-451 were up-regulated by more than two fold,while expression of miR-98,miR-181a,let-7f,let-7g,miR-424 and miR-563 down-regulated by more than two fold in K562/A02 cells.The results of real time RT-PCR were consistent with that of microarray.Of note,differential expressions of miR-451,miR-155.miR-221,let-7f and miR-424 were remarkable.Conclusion K562/A02 cells show a difierent microRNA expression pmfile as compared to its patental K562 cells,suggesting microRNAs including miR-221,miR-155,miR-451,let-7f and miR-424 may be involved in the mechanism of drug resistance in leukemia.These differentially expressed microRNAs provide potential novel targets for overcoming drag-resistance.
