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

发布时间：2018-06-24 08:40

本文选题：miRNA- + Lmo基因　；参考：《中国肿瘤生物治疗杂志》2017年09期

【摘要】：目的:探讨mi R-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mi R-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mi R-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mi R-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mi R-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mi R-223过表达对E6-1细胞成瘤能力的影响。Western blotting检测mi R-223对MAPK信号通路相关蛋白表达的影响。结果:mi R-223在E6-1细胞株中表达水平相对较低(P0.05),双荧光素酶实验证实mi R-223可以直接靶向结合Lmo2的3’UTR区序列。过表达mi R-223后:(1)抑制E6-1细胞的增殖能力(0.16±0.02 vs 1.15±0.21,P0.05);(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,P0.05];(3)调控MAPK信号通路的活性。结论:mi R-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。
[Abstract]:Aim: to investigate the effect of miR-223 on the proliferation and tumorigenesis of (acute lymphoblastic leukemiaall (ALL) E6-1 cells. Methods: the expression of miR-223 in different all cell lines was detected by real-time fluorescent quantitative PCR, and the infection efficiency of E6-1 cell line infected with mi R-223 mimic lentivirus was detected by immunofluorescence staining. The relationship between miR-223 and Lmo2 was detected by double luciferase assay. MTT proliferation assay and clone formation assay were used to detect the effect of overexpression of miR-223 on the proliferation and clone formation of E6-1 cell line. The effect of overexpression of mi R-223 on the tumorigenic ability of E6-1 cells was detected by tumorigenesis assay in nude mice. Western blotting was used to detect the effect of mi R-223 on the expression of MAPK-related proteins. Results the expression level of 1: mi R-223 was relatively low in E6-1 cell line (P0.05). The results of double luciferase assay confirmed that miR-223 could directly target the 3 UTR region of Lmo2. After overexpression of miR-223: (1) inhibition of proliferation of E6-1 cells (0.16 卤0.02 vs 1.15 卤0.21); (_ 2); inhibition of tumorigenic ability of E6-1 cells in nude mice [(0.56 卤0.08) vs (1.69 卤0.22) g P 0.05]; (3) regulation of the activity of MAPKs. Conclusion: 1 / mi R-223 can target Lmo2 to regulate the proliferation, clone formation and tumorigenesis of all cells through MAPK signaling pathway.
【作者单位】：河南大学淮河医院血液科;
【分类号】：R733.71

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