自噬相關基因Beclin1沉默對乳腺癌細胞阿霉素敏感性的影響研究
發(fā)布時間：2019-01-22 18:55
【摘要】：目的研究自噬沉默相關基因Beclin1對乳腺癌MDA-MB-231細胞阿霉素耐藥的影響;方法建立阿霉素耐藥乳腺癌細胞株,起名為MDA-MB-231/ADR,MTT法檢測Beclin1基因沉默對MDA-MB-231/ADR細胞阿霉素化療敏感性的影響,計算IC_(50)值。流式細胞術檢測阿霉素誘導MDA-MB-231/ADR細胞凋亡,Western blot法檢測Bcl-2、Bax、cleaved-caspase3、Beclin1、LC3I/II的表達;結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細胞IC_(50)值(0.26±0.03)μg/ml升高,差異有統(tǒng)計學意義(P0.01)。沉默Beclin1基因增強MDA-MB-231/ADR細胞對阿霉素敏感性,IC_(50)值下降為(2.98±0.58)μg/ml,與si RNA-NC組相比,差異有統(tǒng)計學意義(P0.01)。Beclin1沉默抑制細胞自噬,Beclin1、LC3I/II的表達降低,差異有統(tǒng)計學意義(P0.05)。沉默Beclin1基因增加了阿霉素誘導MDA-MB-231/ADR的細胞凋亡,降低Bcl-2表達,升高Bax、cleaved-caspase3表達;結論 Beclin1基因沉默增加阿霉素耐藥株MDA-MB-231/ADR對阿霉素的敏感性,其可能機制為抑制細胞自噬。
[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】： 三峽大學第一臨床醫(yī)學院;
【基金】：湖北省自然科學基金(2014CFB307)
【分類號】：R737.9
本文編號：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 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】： 三峽大學第一臨床醫(yī)學院;
【基金】：湖北省自然科學基金(2014CFB307)
【分類號】：R737.9
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1 李迎娟;細胞自噬基因Beclin1在細針穿刺乳腺癌組織中的表達及其與p53和Bcl-2的相關性[D];河北醫(yī)科大學;2012年
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