【摘要】：目的升高或者降低信號(hào)轉(zhuǎn)導(dǎo)因子與轉(zhuǎn)錄激活因子3(STAT3)的表達(dá),分析其變化對(duì)于細(xì)胞的增殖分化以及敏感性有何意義。方法構(gòu)建重組質(zhì)粒真核表達(dá)載體siRNASTAT3,將重組表達(dá)質(zhì)粒用脂質(zhì)體Lipofactamine 3000轉(zhuǎn)染人慢性粒細(xì)胞白血病K562細(xì)胞,通過嘌呤霉素持續(xù)單克隆篩選,Western blot法鑒定,進(jìn)而篩選出穩(wěn)定低表達(dá)組(低表達(dá)對(duì)照組、低表達(dá)1組、低表達(dá)2組)和高表達(dá)組(高表達(dá)對(duì)照組、高表達(dá)組)STAT3細(xì)胞株。將正處于增殖周期的細(xì)胞經(jīng)不同濃度的伊馬替尼作用后,MTT法檢測(cè)細(xì)胞生長(zhǎng)抑制率,繪制細(xì)胞增殖曲線。同時(shí)檢測(cè)轉(zhuǎn)染后細(xì)胞株BCR/ABL融合基因相關(guān)蛋白的表達(dá)變化。結(jié)果與低表達(dá)對(duì)照組相比,低表達(dá)STAT3(低表達(dá)1組、低表達(dá)2組)的K562細(xì)胞組使用伊馬替尼治療有著較高的抑制率,在不同濃度作用下,其抑制率差異有統(tǒng)計(jì)學(xué)意義(P0.05)。過表達(dá)STAT3后,與高表達(dá)對(duì)照組相比,在相同濃度梯度伊馬替尼作用下,其抑制率差異有統(tǒng)計(jì)學(xué)意義(P0.05)。低表達(dá)對(duì)照組與高表達(dá)對(duì)照組中伊馬替尼抑制率差異無統(tǒng)計(jì)學(xué)意義。伊馬替尼治療靶向基因BCR/ABL相關(guān)蛋白中的P53、HAUSP和PTEN蛋白表達(dá)趨勢(shì)和STAT3基本一致。結(jié)論低表達(dá)STAT3蛋白可加強(qiáng)K562細(xì)胞對(duì)伊馬替尼的敏感性,抑制STAT3蛋白有望提高伊馬替尼對(duì)慢性粒細(xì)胞白血病的療效。
[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.
【作者單位】： 安徽醫(yī)科大學(xué)第一附屬醫(yī)院血液內(nèi)科;
【基金】：安徽高校省級(jí)自然科學(xué)研究項(xiàng)目(編號(hào):KJ2016A352)
【分類號(hào)】：R733.72

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