miR-223通过Lmo2基因和MAPK信号通路调控急性淋巴细胞白血病的恶性生物学行为

目的:探讨miR-223调控急性淋巴细胞白血病(acute lymphoblastic leukemia,ALL)E6-1细胞的增殖和体内成瘤能力及其作用机制.方法:应用实时荧光定量PCR检测miR-223在不同ALL细胞株中的表达水平,免疫荧光染色法检测携带miR-223mimic慢病毒感染E6-1细胞株的感染效率,双荧光素酶实验检测miR-223和Lmo2的相互结合关系,MTT增殖实验和克隆形成实验检测过表达miR-223对E6-1细胞株增殖和克隆形成能力的影响.裸鼠体内成瘤实验检测miR-223过表达对E6-1细胞成瘤能力的影响.Western blotting检测miR-223对MAPK信号通路相关蛋白表达的影响.结果:miR-223在E6-1细胞株中表达水平相对较低(P＜0.05),双荧光素酶实验证实miR-223可以直接靶向结合Lmo2的3'UTR区序列.过表达miR-223后:(1)抑制E6-1细胞的增殖能力(0.16±0.02 vs 1.15±0.21,P＜0.05);(2)抑制E6-1细胞的裸鼠体内成瘤能力[(0.56±0.08) vs (1.69±0.22)g,P＜0.05];(3)调控MAPK信号通路的活性.结论:miR-223可靶向作用Lmo2通过MAPK信号通路调控ALL细胞的增殖、克隆形成和体内成瘤能力.

miR-223 regulates malignant biological behavior of acute lymphoblastic leukemia cells through targeting Lmo2 gene and MAPK signal pathway

Objective:To investigate the effect and mechanism of miR-223 regulating the proliferation and in vivo tumorigenesis of acute lymphoblastic leukemia (ALL) E6-1 cells.Methods:The expression of miR-223 in different ALL cell lines was detected by Real-time fluorescence quantitative PCR.Immunofluorescence was used to detect the transfection efficiency of miR-223mimic-lenitvirus transfected E6-1 cell line.Double luciferase assay was used to detect the binding between miR-223 and Lmo2.MTT assay and clone formation assay were used to detect the effect of miR-223 on the proliferation and clone formation ability of E6-1 cell line.The effect of miR-223 expression on the tumorigenic ability of E6-1 cells was detected by xenograft formation in nude mice.Western blotting was used to detect the effect of miR-223 on the expressions of MAPK signal pathway related proteins.Results:The expression level ofmiR-223 in E6-1 cell line was relatively low (P＜0.05).Double luciferase assay confirmed that miR223 could directly target the 3'UTR ofLmo2.Overexpression of miR-223 inhibited the proliferation (0.16 ± 0.02 vs 1.15 ± 0.21,P＜0.05) and tumorigenesis ([0.56 ± 0.08]g vs [1.69± 0.22]g,P＜ 0.05) of E6-1 cells and regulated the activity of MAPK signal pathway.Conclusion:miR-223 can regulate the proliferation,clone formation and in vivo tumorigenesis of ALL cells through targeting Lmo2 and MAPK signal pathway.