【摘要】：目的研究自噬沉默相關(guān)基因Beclin1對(duì)乳腺癌MDA-MB-231細(xì)胞阿霉素耐藥的影響;方法建立阿霉素耐藥乳腺癌細(xì)胞株,起名為MDA-MB-231/ADR,MTT法檢測(cè)Beclin1基因沉默對(duì)MDA-MB-231/ADR細(xì)胞阿霉素化療敏感性的影響,計(jì)算IC_(50)值。流式細(xì)胞術(shù)檢測(cè)阿霉素誘導(dǎo)MDA-MB-231/ADR細(xì)胞凋亡,Western blot法檢測(cè)Bcl-2、Bax、cleaved-caspase3、Beclin1、LC3I/II的表達(dá);結(jié)果成功建立阿霉素耐藥株MDA-MB-231/ADR,其IC_(50)值為(8.51±0.96)μg/ml,較MDA-MB-231細(xì)胞IC_(50)值(0.26±0.03)μg/ml升高,差異有統(tǒng)計(jì)學(xué)意義(P0.01)。沉默Beclin1基因增強(qiáng)MDA-MB-231/ADR細(xì)胞對(duì)阿霉素敏感性,IC_(50)值下降為(2.98±0.58)μg/ml,與si RNA-NC組相比,差異有統(tǒng)計(jì)學(xué)意義(P0.01)。Beclin1沉默抑制細(xì)胞自噬,Beclin1、LC3I/II的表達(dá)降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。沉默Beclin1基因增加了阿霉素誘導(dǎo)MDA-MB-231/ADR的細(xì)胞凋亡,降低Bcl-2表達(dá),升高Bax、cleaved-caspase3表達(dá);結(jié)論 Beclin1基因沉默增加阿霉素耐藥株MDA-MB-231/ADR對(duì)阿霉素的敏感性,其可能機(jī)制為抑制細(xì)胞自噬。
[Abstract]:Objective to study the effect of autophagy silencing related gene Beclin1 on adriamycin resistance in MDA-MB-231 cells of breast cancer. Methods the adriamycin resistant breast cancer cell line was established. The effect of Beclin1 gene silencing on the chemosensitivity of MDA-MB-231/ADR cells to doxorubicin was detected by MDA-MB-231/ADR,MTT assay. The value of IC_ (50) was calculated. Flow cytometry was used to detect the expression of Bcl-2,Bax,cleaved-caspase3,Beclin1,LC3I/II in MDA-MB-231/ADR cells induced by doxorubicin. Results the IC_ (50) value of MDA-MB-231/ADR, was (8.51 鹵0.96) 渭 g / ml, which was higher than that of MDA-MB-231 cells (0.26 鹵0.03) 渭 g/ml. The difference was statistically significant (P0.01). Silencing Beclin1 gene enhanced the sensitivity of MDA-MB-231/ADR cells to adriamycin, and IC_ (50) decreased to (2.98 鹵0.58) 渭 g / ml, which was significantly different from that in si RNA-NC group (P0.01). Beclin1 silencing inhibited autophagy. The expression of Beclin1,LC3I/II decreased, the difference was statistically significant (P0.05). Silencing Beclin1 gene increased the apoptosis of MDA-MB-231/ADR cells induced by doxorubicin, decreased the expression of Bcl-2 and increased the expression of Bax,cleaved-caspase3. Conclusion Beclin1 gene silencing increases the sensitivity of adriamycin-resistant MDA-MB-231/ADR to adriamycin by inhibiting autophagy.
【作者單位】： 三峽大學(xué)第一臨床醫(yī)學(xué)院;
【基金】：湖北省自然科學(xué)基金(2014CFB307)
【分類號(hào)】：R737.9

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