STAT3蛋白水平表达的高低对慢性粒细胞白血病伊马替尼治疗敏感性的研究

发布时间：2018-05-30 08:05

本文选题：STAT3 + 慢性粒细胞白血病　；参考：《安徽医科大学》2017年硕士论文

【摘要】：目的:升高或者降低K562细胞株中信号转导因子与转录激活因子3(Signal transducer and activator of transcription,STAT3)的表达,分析其变化对于CML细胞的增殖分化及伊马替尼的治疗敏感性有何意义。方法:将三组低表达和两组高表达的重组质粒真核表达载体si RNA-STAT3用脂质体Lipofactamin 3000转染人慢性粒细胞白血病K562细胞株,通过嘌呤霉素持续单克隆筛选,Western-blot印迹法鉴定,进而筛选出稳定低表达组(sh-vector、sh STAT3-1、sh STAT3-2)和高表达组(Overexpression vector、Overexpression)STAT3细胞株。将处于增殖周期的细胞经不同浓度的伊马替尼(imatinib,IM)作用后,MTT法检测细胞生长抑制率,绘制细胞增殖曲线。同时Western-blot法检测转染后细胞株BCR/ABL相关蛋白P53、HAUSP和PTEN的表达变化。结果:与其对照组(sh-vector)相比,低表达STAT3蛋白(sh STAT3-1、sh STAT3-2)的K562细胞组使用IM治疗有着较高的抑制率,在0.1μM、0.25μM、0.5μM和1.0μM作用下,其抑制率达到了(76.3±1.84)%、(80.4±1.90)%、(84.4±2.61)%和(94.3±0.19)%,且差异均有统计学意义(p0.05)。提高STAT3(Overexpression)蛋白表达水平后,跟其对照组(Overexpression vector)相比,在相同浓度梯度IM作用下,其抑制率在两组平均下降到(58.2±1.27)%、(63.2±2.32)%、(67.9±1.00)%和(74.1±0.76)%,且差异均有统计学意义(p0.05)。两对照组中IM抑制率差异无统计学意义(p0.05)。IM治疗靶向基因酪氨酸激酶BCR-ABL相关蛋白表达中,P53、HAUSP和PTEN蛋白表达趋势和STAT3基本一致,说明STAT3在K562细胞株中的表达有意义。结论:低表达STAT3蛋白可提高K562细胞对IM的敏感性,抑制STAT3蛋白有望提高IM对CML的疗效。
[Abstract]:Aim: to increase or decrease the expression of signal transduction factor (3(Signal transducer and activator of) and activator 3(Signal transducer and activator of transcription3 (STAT3) in K562 cell line and analyze the significance of these changes in the proliferation and differentiation of CML cells and the therapeutic sensitivity of imatinib. Methods: three groups of low expression and two groups of high expression eukaryotic expression vector si RNA-STAT3 were transfected into human chronic myeloid leukemia K562 cell line with liposome Lipofactamin 3000 and identified by purine mycin continuous monoclonal screening and Western-blot blotting. Furthermore, stable low expression group STAT3-1nsh STAT3-2 and high expression group Overexpression vector expression STAT3 cell line were screened. Cell growth inhibition rate was measured by MTT assay and cell proliferation curve was plotted after different concentrations of imatinib (imatinib) were added to the cells in the proliferative cycle. At the same time, the expression of BCR/ABL associated protein P53, HAUSP and PTEN were detected by Western-blot. Results: compared with the control group, K562 cells with low expression of STAT3 protein stash STAT3-1sh STAT3-2) had a higher inhibitory rate after treatment with IM. The inhibitory rates reached 76.3 卤1.844.40 卤1.90,84.4 卤2.61% and 94.3 卤0.19m respectively under the treatment of 0.1 渭 M 0.25 渭 M and 1.0 渭 M, and the difference was statistically significant. After increasing the expression level of STAT3Overexpressionprotein, compared with the control group, under the same concentration gradient IM, the inhibition rate of STAT3Overexpressionprotein decreased to 58.2 卤1.27g + 67.9 卤1.00% and 74.1 卤0.76% respectively, and the difference was statistically significant (p 0.05). There was no significant difference in the inhibition rate of IM between the two control groups. There was no statistical significance in the expression of BCR-ABL related protein of tyrosine kinase targeting gene in IM. The expression trend of P53HAP and PTEN protein in K562 cell line was similar to that of STAT3, which indicated that the expression of STAT3 in K562 cell line was significant. Conclusion: the low expression of STAT3 protein can increase the sensitivity of K562 cells to IM, and the inhibition of STAT3 protein may improve the therapeutic effect of IM on CML.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R733.72

【参考文献】