长链非编码RNA HIT对急性淋巴细胞白血病SUP-B15细胞系伊马替尼抵抗的影响

Effect of long non-coding RNA HIT on imatinib resistance in acute lymphoblastic leukemia SUP-B15 cell line

目的探究长链非编码RNA(LncRNA)HIT对急性淋巴细胞白血病(ALL)SUP-B15细胞系伊马替尼(IM)抵抗的影响及相关机制。方法浓度梯度IM及生理盐水处理SUP-B15细胞作为IMR组及Control组,慢病毒转染LncRNA HIT shRNA1#及2#载体于IMR组细胞作为IMR shHIT1#及2#组细胞。CCK-8实验检测细胞IM半抑制浓度(IC 50),荧光定量PCR技术(qPCR)检测细胞HIT、QKI、Oct4及SOX2 mRNA水平,蛋白免疫印迹实验检测细胞QKI、Oct4及SOX2蛋白表达水平。结果相比Control细胞,IMR、IMR shHIT1#及2#细胞中IM IC 50明显增高,LncRNA HIT表达、Oct4及Sox2的mRNA及蛋白表达明显上调,QKI mRNA及蛋白表达明显下调;相比IMR细胞,IMR shHIT1#及2#细胞IM IC 50明显降低,LncRNA HIT表达、Oct4及Sox2 mRNA及蛋白表达明显下调,QKI mRNA及蛋白表达明显上调,差异均具有统计学意义(P<0.05)。结论LncRNA HIT可通过抑制QKI蛋白的表达进而上调Sox2及Oct4,介导ALL细胞IM抵抗的形成。

Aim To investigate the effect of long non-coding RNA(LncRNA)HIT on the resistance of imatinib(IM)in SUP-B15 cell line of acute lymphoblastic leukemia(ALL)and the related mechanisms.Methods SUP-B15 cells were treated with concentration gradient IM and saline as IMR and control groups.The lentivirus transfected LncRNA HIT shRNA1#and 2#vector in IMR group cells to knock down the HIT expression as IMR shHIT1#and 2#group cells.CCK-8 assay was used to detect IM half-inhibitory concentration(IC 50).Fluorescence quantitative PCR(qPCR)was applied to detect the expression levels of LncRNA HIT,QKI,Oct4 and Sox2 mRNA.Western blot was employed to detect the expression levels of QKI,Oct4 and Sox2 protein.Results Compared with those in control cells,there was significantly higher IM IC 50,higher expression of LncRNA HIT,Oct4 and Sox2,and lower expression of QKI in groups of IMR,IMR shHIT1#and 2#cells.Compared with those in IMR cells,there was significantly lower IM IC 50,lower expression of LncRNA HIT,Oct4 and Sox2,and higher expression of QKI in groups of IMR shHIT1#and 2#cells.The difference was statistically significant(P<0.05).Conclusions LncRNA HIT can increase the expression of Sox2 and Oct4 via inhibiting the expression of QKI protein,and mediating the formation of IM resistance in ALL cells.