本文選題：白血病　切入點：髓系　出處：《青島大學》2017年碩士論文

【摘要】：目的 研究SOX4基因在慢性髓系白血病(CML)中的作用機制,探討SOX4基因作為CML靶點的可能性。方法 培養(yǎng)慢性髓系白血病K562細胞,將構建好的si RNA-SOX4表達質(zhì)粒p LVE-1680及空載體p LVE經(jīng)脂質(zhì)體轉(zhuǎn)入K562細胞后檢測SOX4 m RNA和蛋白表達水平;q RT-PCR方法檢測p53、bax、c-myc、cyclin D1、bcl-2基因的表達情況;RT-PCR方法檢測Foxo3a、C/EBPα基因的表達情況;CCK8法檢測細胞增殖情況;基因芯片檢測干擾組和對照組中差異基因的表達。結(jié)果 RT-PCR和Western blot檢測結(jié)果顯示含目的基因si RNA-SOX4的K562細胞株已成功構建表達;si RNA SOX4-K562組C/EBPα、bax、Foxo3a、cyclin D1基因的m RNA表達量分別為0.1609±0.0095、1.2410±0.0602、1.1450±0.0461、1.7240±0.0657,顯著高于K562組及K562-空載體組(p0.01,p0.05,p0.05,p0.05);c-myc、bcl-2、p53基因m RNA表達量分別為0.9171±0.00953、0.6507±0.0090、0.4952±0.0220,顯著低于K562組及K562-空載體組(p0.01,p0.01,p0.01);結(jié)果顯示si RNASOX4-K562組細胞增殖程度顯著低于K562細胞組及K562-空載體細胞組(p0.05);以差異基因2倍為限,基因芯片檢測結(jié)果干擾組較對照組升高的基因有4個,較對照組降低的基因有7個。兩組差異表達基因Pathyway富集功能分析結(jié)果顯示最集中的pathyway是血小板激活、白細胞跨膜遷移以及病灶粘連。結(jié)論 SOX4基因在慢性髓系白血病中發(fā)揮癌基因的作用,在K562細胞中干擾SOX4能夠抑制細胞增殖并促進細胞凋亡;基因芯片檢測結(jié)果表明SOX4可能通過下調(diào)RAP1A,上調(diào)SNX2、ZNF486、AX747507等基因表達水平來干擾CML的發(fā)病過程。上述結(jié)果顯示干擾SOX4基因的表達可能是慢性髓系白血病治療的潛在靶點。
[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 偽 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 偽 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 鹵0.009530.6507 鹵0.00900.4952 鹵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 鹵0.009530.6507 鹵0.0090) 0.4952 鹵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.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R733.72

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本文編號：1670775