STAT3蛋白表达水平的高低对慢性粒细胞白血病伊马替尼治疗敏感性的影响

发布时间：2018-09-08 17:26

【摘要】：目的升高或者降低信号转导因子与转录激活因子3(STAT3)的表达,分析其变化对于细胞的增殖分化以及敏感性有何意义。方法构建重组质粒真核表达载体siRNASTAT3,将重组表达质粒用脂质体Lipofactamine 3000转染人慢性粒细胞白血病K562细胞,通过嘌呤霉素持续单克隆筛选,Western blot法鉴定,进而筛选出稳定低表达组(低表达对照组、低表达1组、低表达2组)和高表达组(高表达对照组、高表达组)STAT3细胞株。将正处于增殖周期的细胞经不同浓度的伊马替尼作用后,MTT法检测细胞生长抑制率,绘制细胞增殖曲线。同时检测转染后细胞株BCR/ABL融合基因相关蛋白的表达变化。结果与低表达对照组相比,低表达STAT3(低表达1组、低表达2组)的K562细胞组使用伊马替尼治疗有着较高的抑制率,在不同浓度作用下,其抑制率差异有统计学意义(P0.05)。过表达STAT3后,与高表达对照组相比,在相同浓度梯度伊马替尼作用下,其抑制率差异有统计学意义(P0.05)。低表达对照组与高表达对照组中伊马替尼抑制率差异无统计学意义。伊马替尼治疗靶向基因BCR/ABL相关蛋白中的P53、HAUSP和PTEN蛋白表达趋势和STAT3基本一致。结论低表达STAT3蛋白可加强K562细胞对伊马替尼的敏感性,抑制STAT3蛋白有望提高伊马替尼对慢性粒细胞白血病的疗效。
[Abstract]:Objective to increase or decrease the expression of signal transduction factor and activator of transcription 3 (STAT3), and to analyze the significance of the changes in the expression of signal transduction factor and activator 3 (STAT3) for cell proliferation, differentiation and sensitivity. Methods Recombinant eukaryotic expression vector siRNASTAT3, was constructed and transfected into human chronic myeloid leukemia K562 cells by liposome Lipofactamine 3000. The recombinant plasmid was identified by purine mycin continuous monoclonal screening and Western blot. The STAT3 cell lines of stable and low expression group (low expression control group, low expression group 1, low expression group 2) and high expression group (high expression control group, high expression group) were screened out. Cell growth inhibition rate was measured by MTT assay and cell proliferation curve was plotted after different concentrations of imatinib were treated with imatinib. At the same time, the expression of BCR/ABL fusion gene related protein was detected after transfection. Results compared with the low expression control group, the K562 cell group with low expression of STAT3 (low expression group 1, low expression group 2) had a higher inhibitory rate when treated with imatinib, and there was significant difference in the inhibition rate under different concentrations (P0.05). After overexpression of STAT3, there was significant difference in inhibition rate between the two groups under the same concentration gradient imatinib (P0.05). There was no significant difference in the inhibition rate of imatinib between low expression control group and high expression control group. The expression trend of P53-HAUSP and PTEN in imatinib targeted gene BCR/ABL related protein was basically consistent with that of STAT3. Conclusion the low expression of STAT3 protein can enhance the sensitivity of K562 cells to imatinib, and the inhibition of STAT3 protein may improve the therapeutic effect of imatinib on chronic myeloid leukemia.
【作者单位】：安徽医科大学第一附属医院血液内科;
【基金】：安徽高校省级自然科学研究项目(编号:KJ2016A352)
【分类号】：R733.72

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