SOX4基因在慢性髓系白血病中的机制研究

发布时间：2018-03-27 08:31

本文选题：白血病　切入点：髓系　出处：《青岛大学》2017年硕士论文

【摘要】：目的研究SOX4基因在慢性髓系白血病(CML)中的作用机制,探讨SOX4基因作为CML靶点的可能性。方法培养慢性髓系白血病K562细胞,将构建好的si RNA-SOX4表达质粒p LVE-1680及空载体p LVE经脂质体转入K562细胞后检测SOX4 m RNA和蛋白表达水平;q RT-PCR方法检测p53、bax、c-myc、cyclin D1、bcl-2基因的表达情况;RT-PCR方法检测Foxo3a、C/EBPα基因的表达情况;CCK8法检测细胞增殖情况;基因芯片检测干扰组和对照组中差异基因的表达。结果 RT-PCR和Western blot检测结果显示含目的基因si RNA-SOX4的K562细胞株已成功构建表达;si RNA SOX4-K562组C/EBPα、bax、Foxo3a、cyclin D1基因的m RNA表达量分别为0.1609±0.0095、1.2410±0.0602、1.1450±0.0461、1.7240±0.0657,显著高于K562组及K562-空载体组(p0.01,p0.05,p0.05,p0.05);c-myc、bcl-2、p53基因m RNA表达量分别为0.9171±0.00953、0.6507±0.0090、0.4952±0.0220,显著低于K562组及K562-空载体组(p0.01,p0.01,p0.01);结果显示si RNASOX4-K562组细胞增殖程度显著低于K562细胞组及K562-空载体细胞组(p0.05);以差异基因2倍为限,基因芯片检测结果干扰组较对照组升高的基因有4个,较对照组降低的基因有7个。两组差异表达基因Pathyway富集功能分析结果显示最集中的pathyway是血小板激活、白细胞跨膜迁移以及病灶粘连。结论 SOX4基因在慢性髓系白血病中发挥癌基因的作用,在K562细胞中干扰SOX4能够抑制细胞增殖并促进细胞凋亡;基因芯片检测结果表明SOX4可能通过下调RAP1A,上调SNX2、ZNF486、AX747507等基因表达水平来干扰CML的发病过程。上述结果显示干扰SOX4基因的表达可能是慢性髓系白血病治疗的潜在靶点。
[Abstract]:Objective to study the mechanism of SOX4 gene in chronic myeloid leukemia (CML) and explore the possibility of SOX4 gene as a target of CML. Expression level of SOX4 m RNA and protein expression in K562 cells were detected by using the constructed si RNA-SOX4 expression plasmid p LVE-1680 and empty vector p LVE after transfection into K562 cells by liposome. The expression of bcl-2 gene was detected by p53 BaxCX c-myctrin cyclin D1 BCL 2 gene expression. RT-PCR method was used to detect the expression of Foxo3aAU C% EBP 伪 gene in K562 cells. Cell proliferation was detected by HPLC. Results the results of RT-PCR and Western blot analysis showed that K562 cell line containing the target gene si RNA-SOX4 had been successfully constructed for the expression of RNA in C/EBP 伪 Foxo3a cyclin D1 gene in the RNA SOX4-K562 group. The expression of p53 gene m RNA in K562 group and K562-empty carrier group was significantly higher than that in K562 group and K562-empty vector group, the expression of p53 gene m RNA was 0.9171 卤0.009530.6507 卤0.00900.4952 卤0.0220, which was significantly lower than that in K562 group and K562-empty vector group, the results showed that the proliferation of p0.01p0.01p0.01p0.01p0.01in RNASOX4-K562 group was significantly lower than that in K562 cell group and K562-empty carrier group (0.9171 卤0.009530.6507 卤0.0090) 0.4952 卤0.0220. the results showed that the proliferative degree of p0.01p0.01p0.01p0.01in RNASOX4-K562 group was significantly lower than that in K562 cell group and K562-empty carrier group, and that in K562-empty vector group was significantly lower than that in K562 cell group and K562-empty carrier group. The expression of p53 gene m RNA in K562 group was significantly lower than that in K562 group and K562-empty vector group. The difference gene is limited to 2 times, The results of microarray analysis showed that 4 genes were higher in interference group than those in control group, and 7 genes were lower than those in control group. The results of Pathyway enrichment analysis of differentially expressed genes in two groups showed that platelet activation was the most concentrated pathyway. Conclusion SOX4 gene plays the role of oncogene in chronic myeloid leukemia and interfering with SOX4 in K562 cells can inhibit cell proliferation and promote cell apoptosis. The results of microarray analysis suggested that SOX4 might interfere with the pathogenesis of CML by down-regulating RAP1A and up-regulating the expression level of SNX2ANF486AX747507, which suggested that interfering with the expression of SOX4 gene might be a potential target for the treatment of chronic myeloid leukemia.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R733.72

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