摘要
目的:观察长链非编码-HOX反义基因间RNA髓样1(LncRNA-HOTAIRM1)下调对人白血病T淋巴细胞Jurkat增殖及凋亡的影响,并初步探讨其作用机理。方法:体外培养Jurkat细胞,随机分为对照组、HOTAIRM1siRNA-NC组、HOTAIRM1 siRNA组;利用实时荧光定量PCR法检测转染后各组Jurkat细胞LncRNA-HOTAIRM1mRNA、KIT受体酪氨酸激酶(KIT) mRNA、丝氨酸-苏氨酸激酶(AKT) mRNA的表达情况;采用CCK-8法检测各组Jurkat细胞的增殖情况;利用Annexin V-FITC/PI双染法检测各组Jurkat细胞的凋亡情况;采用蛋白印迹分析法检测KIT、AKT、p-KIT、p-AKT、B淋巴细胞瘤-2基因(BCL-2)及半胱氨酸天冬氨酸酶3(Caspase-3)蛋白表达的情况。结果:与对照组和HOTAIRM1 siRNA-NC组相比,HOTAIRM1 siRNA组Jurkat细胞LncRNA-HOTAIRM1 mRNA表达水平、细胞存活率、KIT mRNA、AKT mRNA、p-KIT、p-AKT及BCL-2蛋白表达水平均显著降低(P<0.05),Cleared Caspase-3蛋白表达水平、Jurkat细胞凋亡率显著升高(P<0.05)。结论:LncRNA-HOTAIRM1可能通过KIT/AKT信号通路,抑制Jurkat细胞增殖并诱导其凋亡。
Objective: To observe the effects of down-regulation of long non-coding RNA HOX antisense intergenic RNA myeloid 1(LncRNA-HOTAIRM1) to the proliferation and apoptosis of Jurkat in human leukemia T lymphocytes,and explore its mechanism. Methods: Jurkat cells were cultured in vitro and randomly divided into control group,HOTAIRM1 siRNA-NC group and HOTAIRM1 siRNA group;the expressions of LncRNA-HOTAIRM1 mRNA, KIT receptor tyrosine kinase(KIT) mRNA and serine threonine kinase(AKT) mRNA in Jurkat cells were detected by realtime fluorescence quantification(RT-qPCR);the proliferation of Jurkat cells in each groups was detected by CCK-8 method;the apoptosis of Jurkat cells in each groups was detected by Annexin V-FITC/PI double staining;the expressions of KIT, AKT, p-KIT, p-AKT, B-lymphoma-2 gene(BCL-2) and Caspase-3 were detected by Western blot. Results:Compared with the cells in the control group and HOTAIRM1 siRNA-NC group, the expression level of LncRNAHOTAIRM1 mRNA, cell survival rate, expression levels of KIT mRNA, AKT mRNA, p-KIT, p-AKT and BCL-2 proteins in Jurkat cells in HOTAIRM1 siRNA group were significantly lower(P<0.05), while the expression level of Cleared Caspase-3 protein and Jurkat cell apoptosis rate were significantly higher(P<0.05). Conclusion: LncRNAHOTAIRM1 may inhibit Jurkat cell proliferation and induce apoptosis through KIT/AKT signaling pathway.
作者
李玉茹
燕文敬
蔡力力
邓新立
LI Yu-Ru;YAN Wen-Jing;CAI Li-Li;DENG Xin-Li(Department of Laboratory Medicine,Second Medical Center of PLA General Hospital,Beijing 100853,China)
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2021年第4期1123-1128,共6页
Journal of Experimental Hematology
