微小RNA-153靶向磷脂酰肌醇3激酶/丝氨酸-苏氨酸蛋白激酶通路调控急性淋巴细胞白血病细胞增殖

摘要

Objective To investigate the effect of microRNA (miRNA,miR)-153 on the proliferation and apoptosis of in human acute lymphoblastic leukemia (ALL) cell line Jurkat cells by phosphatidylinositol 3 kinase (PI3K)/serine-threonine protein kinase (Akt) pathway.Methods After miR-153 mimics (miR-153 mimics group) and miR-153 inhibitor (miR-153 inhibitor group) were transfected into Jurkat cells,cell proliferation,cycle and apoptosis.was detected;the expression of miR-153,PI3K and Akt mRNA and protein was measured;whether PI3K was the target gene of miR-153.was confirmed.Results Compared with the negative control group,the expression of miR-153 in the miR-153 mimics group was significantly increased by approximately 10-fold (t =39.596,P ＜ 0.01),and the expression of miR-153 was significantly decreased in the miR-153 inhibitor group (t =14.651,P ＜ 0.01).The activity of the cells in the miR-153 mimics group was significantly decreased (t =10.931,P ＜ 0.01),and the activity of the miR-153 inhibitor group was significantly increased (t =7.640,P ＜ 0.01).The expression of PI3K total protein,PI3K mRNA and p-Akt in the miR-153 mimics group was significantly decreased (t =7.161,P ＜ 0.01;t =6.684,P ＜ 0.01;t =10.769,P ＜ 0.01),and the expression of PI3K total protein,PI3K mRNA and p-Akt in the miR-153 inhibitor group was significantly increased (t =3.998,P ＜ 0.05;t =8.902,P ＜ 0.01;t =7.078,P ＜ 0.01).The proportion of G0/G1 phase cells increased significantly,while the S phase and G2/M phasecells decreased significantly (t =3.786,P＜0.05;t =3.012,P＜0.05;t =4.948,P＜0.01) in the miR-153 mimics group;the G0/G1 phase cells decreased significantly,and the proportion of cells in S phase and G2/M phase increased significantly (t =5.356,P ＜0.01;t =3.055,P ＜0.05;t =3.893,P ＜0.05) in the miR-153 inhibitor group.The apoptosis rate of the miR-153 mimics group was significantly increased (t =4.673,P ＜0.05),while the apoptotic rate of the miR-153 inhibitor group was significant decline (t =5.618,P ＜0.01).Compared to the empty plasmid group,the relative luciferase activity of 3'untranslated regions (3'UTR) plasmid group was significantly reduced (t =7.925,P ＜ 0.01).Conclusion MiR-153 can directly target PI3K/Akt signaling pathway and regulate Jurkat cell proliferation and apoptosis.%目的 探讨微小RNA(miRNA,miR)-153靶向调控磷脂酰肌醇3激酶(PI3K)/丝氨酸-苏氨酸蛋白激酶(Akt)通路对人急性淋巴细胞白血病(ALL)细胞Jurkat的影响.方法 miR-153模拟物(miR-153模拟物组)、miR-153抑制物(miR-153抑制物组)转染Jurkat细胞后,观察细胞增殖、周期和凋亡,检测miR-153、PI3K和Akt mRNA及蛋白表达,并验证PI3K是否为miR-153的靶基因.结果 与阴性对照组比较,miR-153模拟物组miR-153表达显著升高约10倍(t=39.596,P＜0.01),miR-153抑制物组miR-153表达明显降低(t=14.651,P＜0.01).miR-153模拟物组细胞活性显著下降(t=10.931,P＜0.01),miR-153抑制物组细胞活性明显升高(t=7.640,P＜0.01).miR-153模拟物组中PI3K总蛋白、PDK mRNA及p-Akt表达明显下降(t=7.161,P＜0.01;t=6.684,P＜0.01;t=10.769,P＜0.01),而miR-153抑制物组PI3K总蛋白、PI3K mRNA及p-Akt表达显著升高(t =3.998,P＜0.05;t=8.902,P＜0.01;t =7.078,P＜0.01).miR-153模拟物组G0/G1期细胞比例明显增多,而S期和G2/M期细胞显著下降(t=3.786,P＜0.05;t =3.012,P＜0.05;t=4.948,P＜0.01);miR-153抑制物组G0/G1期细胞显著下降,S期和G2/M期细胞所占比例明显增加(t =5.356,P＜0.01;t=3.055,P＜0.05;=3.893,P＜0.05).miR-153模拟物组细胞凋亡率明显增加(t =4.673,P＜0.05),而miR-153抑制物组凋亡率显著下降(t=5.618,P＜0.01).与空载质粒组比较,3'端非编码区(3'UTR)质粒组相对荧光素酶活性显著降低(t=7.925,P＜0.01).结论 miR-153能够直接靶向PI3K/Akt信号通路,调控Jurkat细胞增殖和凋亡.