自噬相关基因Beclin1沉默对乳腺癌细胞阿霉素敏感性的影响研究
发布时间：2019-01-22 18:55
【摘要】：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(P0.01)。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(P0.01)。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(P0.05)。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。
[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 卤0.96) 渭 g / ml, which was higher than that of MDA-MB-231 cells (0.26 卤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 卤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.
【作者单位】： 三峡大学第一临床医学院;
【基金】：湖北省自然科学基金(2014CFB307)
【分类号】：R737.9
本文编号：2413470
[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 卤0.96) 渭 g / ml, which was higher than that of MDA-MB-231 cells (0.26 卤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 卤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.
【作者单位】： 三峡大学第一临床医学院;
【基金】：湖北省自然科学基金(2014CFB307)
【分类号】：R737.9
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1 李迎娟;细胞自噬基因Beclin1在细针穿刺乳腺癌组织中的表达及其与p53和Bcl-2的相关性[D];河北医科大学;2012年
，本文编号：2413470
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