本文選題：STAT3 + 慢性粒細(xì)胞白血病��； 參考：《安徽醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:升高或者降低K562細(xì)胞株中信號(hào)轉(zhuǎn)導(dǎo)因子與轉(zhuǎn)錄激活因子3(Signal transducer and activator of transcription,STAT3)的表達(dá),分析其變化對(duì)于CML細(xì)胞的增殖分化及伊馬替尼的治療敏感性有何意義。方法:將三組低表達(dá)和兩組高表達(dá)的重組質(zhì)粒真核表達(dá)載體si RNA-STAT3用脂質(zhì)體Lipofactamin 3000轉(zhuǎn)染人慢性粒細(xì)胞白血病K562細(xì)胞株,通過(guò)嘌呤霉素持續(xù)單克隆篩選,Western-blot印跡法鑒定,進(jìn)而篩選出穩(wěn)定低表達(dá)組(sh-vector、sh STAT3-1、sh STAT3-2)和高表達(dá)組(Overexpression vector、Overexpression)STAT3細(xì)胞株。將處于增殖周期的細(xì)胞經(jīng)不同濃度的伊馬替尼(imatinib,IM)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)Western-blot法檢測(cè)轉(zhuǎn)染后細(xì)胞株BCR/ABL相關(guān)蛋白P53、HAUSP和PTEN的表達(dá)變化。結(jié)果:與其對(duì)照組(sh-vector)相比,低表達(dá)STAT3蛋白(sh STAT3-1、sh STAT3-2)的K562細(xì)胞組使用IM治療有著較高的抑制率,在0.1μM、0.25μM、0.5μM和1.0μM作用下,其抑制率達(dá)到了(76.3±1.84)%、(80.4±1.90)%、(84.4±2.61)%和(94.3±0.19)%,且差異均有統(tǒng)計(jì)學(xué)意義(p0.05)。提高STAT3(Overexpression)蛋白表達(dá)水平后,跟其對(duì)照組(Overexpression vector)相比,在相同濃度梯度IM作用下,其抑制率在兩組平均下降到(58.2±1.27)%、(63.2±2.32)%、(67.9±1.00)%和(74.1±0.76)%,且差異均有統(tǒng)計(jì)學(xué)意義(p0.05)。兩對(duì)照組中IM抑制率差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)。IM治療靶向基因酪氨酸激酶BCR-ABL相關(guān)蛋白表達(dá)中,P53、HAUSP和PTEN蛋白表達(dá)趨勢(shì)和STAT3基本一致,說(shuō)明STAT3在K562細(xì)胞株中的表達(dá)有意義。結(jié)論:低表達(dá)STAT3蛋白可提高K562細(xì)胞對(duì)IM的敏感性,抑制STAT3蛋白有望提高IM對(duì)CML的療效。
[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 鹵1.844.40 鹵1.90,84.4 鹵2.61% and 94.3 鹵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 鹵1.27g + 67.9 鹵1.00% and 74.1 鹵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.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R733.72

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