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目的研究自噬沉默相关基因Beclin1对乳腺癌MDA-MB-231细胞阿霉素耐药的影响;方法建立阿霉素耐药乳腺癌细胞株,起名为MDA-MB-231/ADR,MTT法检测Beclin1基因沉默对MDA-MB-231/ADR细胞阿霉素化疗敏感性的影响,计算IC_(50)值。流式细胞术检测阿霉素诱导MDA-MB-231/ADR细胞凋亡,Western blot法检测Bcl-2、Bax、cleaved-caspase3、Beclin1、LC3I/II的表达;结果成功建立阿霉素耐药株MDA-MB-231/ADR,其IC_(50)值为(8.51±0.96)μg/ml,较MDA-MB-231细胞IC_(50)值(0.26±0.03)μg/ml升高,差异有统计学意义(P<0.01)。沉默Beclin1基因增强MDA-MB-231/ADR细胞对阿霉素敏感性,IC_(50)值下降为(2.98±0.58)μg/ml,与si RNA-NC组相比,差异有统计学意义(P<0.01)。Beclin1沉默抑制细胞自噬,Beclin1、LC3I/II的表达降低,差异有统计学意义(P<0.05)。沉默Beclin1基因增加了阿霉素诱导MDA-MB-231/ADR的细胞凋亡,降低Bcl-2表达,升高Bax、cleaved-caspase3表达;结论 Beclin1基因沉默增加阿霉素耐药株MDA-MB-231/ADR对阿霉素的敏感性,其可能机制为抑制细胞自噬。
Objective To study the effect of autophagy-related gene Beclin1 on doxorubicin resistance in breast cancer MDA-MB-231 cells.Methods To establish a doxorubicin-resistant breast cancer cell line named MDA-MB-231 / ADR and MTT assay The effect of Beclin1 gene silencing on the chemosensitivity of doxorubicin to MDA-MB-231 / ADR cells was detected and IC 50 values were calculated. Flow cytometry was used to detect the apoptosis of MDA-MB-231 / ADR cells induced by doxorubicin. Western blot was used to detect the expression of Bcl-2, Bax, cleaved-caspase3, Beclin1 and LC3I / The IC50 of strain MDA-MB-231 / ADR was (8.51 ± 0.96) μg / ml, which was higher than that of MDA-MB-231 cells (0.26 ± 0.03) μg / ml Statistical significance (P <0.01). The silencing of Beclin1 enhanced the sensitivity to doxorubicin in MDA-MB-231 / ADR cells, and the IC 50 value decreased to (2.98 ± 0.58) μg / ml, which was significantly different from that in si RNA-NC group P <0.01). Beclin1 silenced the autophagy, and the expression of Beclin1 and LC3I / II decreased, with statistical significance (P <0.05). The silencing of Beclin1 gene increased the apoptosis of MDA-MB-231 / ADR induced by doxorubicin, decreased the expression of Bcl-2 and increased the expression of Bax and cleaved-caspase3. Conclusion Beclin1 silencing increased the expression of doxorubicin-resistant MDA- -231 / ADR on doxorubicin sensitivity, its possible mechanism is to inhibit cell autophagy.