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在全球范围内,乳腺癌是女性癌症死亡的主要原因,2018年约有210万女性疑似患有乳腺癌,占女性癌症的25%[1]。在我国,乳腺癌在女性肿瘤中发病率居首位,死亡率居第5位,近几十年来乳腺癌负担迅速增长[2]。目前,雌激素敏感的乳腺癌患者主要以内分泌治疗为主,然而,缺乏激素受体的乳腺癌细胞通常使用化疗药物,如紫杉醇和阿霉素[3]。但是,化疗药物对肿瘤细胞和正常细胞均表现出毒性反应,这限制了其临床应用。此外,细胞毒等抗肿瘤药物表现出的细胞耐药性进一步限制其应用。因此,迫切需要寻找毒性较小、效果较好的治疗药物。
五味子乙素(schisandrin B, Sch B)是从五味子中提取的主要活性成分[4]。据报道[5],Sch B可通过抑制NF-κB激活及MAPK/ Erk/p38/c-Jnk信号通路激活而有效减轻炎症反应,NASSER等[6]发现,Sch B可通过抑制PI3K/AKT和STA3/JAK2信号通路磷酸化,从而降低细胞内ROS的产生发挥抗前列腺癌作用,Wang等[7]发现,Sch B可通过TGF-b信号通路靶向miR-101-5p抑制大鼠肝纤维化。Dai等[8]发现,Sch B可通过STAT3的磷酸化和核转位发挥抗乳腺癌活性,但是Sch B 如何影响乳腺癌细胞增殖凋亡能力及具体机制尚不清楚。本研究以Sch B为研究对象,探讨其对人乳腺癌MDA-MB-231细胞凋亡的影响及其作用机制。
Schisandrin B induces apoptosis of human breast cancer MDA-MB-231 cells through ROS mediated endoplasmic reticulum stress
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摘要:
目的 研究五味子乙素对人乳腺癌MDA-MB-231细胞凋亡的影响及其作用机制。 方法 用细胞计数试剂(CCK-8)检测不同浓度五味子乙素对MDA-MB-231细胞存活率的影响;五味子乙素(10、20、40 μmol/L)作用 MDA-MB-231 细胞 24 h,分别用Annexin V-FITC/PI检测细胞凋亡情况;用DCFA-DA荧光探针检测细胞内活性氧(ROS)水平;用Western blot法检测细胞凋亡及内质网应激相关蛋白(Bcl-2、Bax、CHOP、GPR78、PERK、p-PERK、p-eIF2α、eIF2)的表达。 结果 与空白组比较,随着五味子乙素浓度增大,细胞存活率明显降低,其IC50为19.16 μmol/L;与对照组比较,五味子乙素(10、20、40 μmol/L)均能抑制细胞克隆形成(P<0.05),且呈剂量依赖;五味子乙素(10、20、40 μmol/L)均可诱导细胞凋亡(P<0.05),使抗凋亡蛋白BCL-2的表达显著降低,促凋亡蛋白Bax的表达显著升高(P<0.05);五味子乙素(10、20、40 μmol/L)显著升高细胞内ROS水平(P<0.05),且呈剂量依赖;五味子乙素(10、20、40 μmol/L)能够激发内质网应激,使内质网应激相关蛋白CHOP、GPR78、p-eIF2α表达增多(P<0.05),且呈剂量依赖。 结论 五味子乙素可能通过ROS介导内质网应激诱导MDA-MB-231细胞凋亡。 Abstract:Objective To study the effects of schisandrin B (Sch B) on the apoptosis of human breast cancer MDA-MB-231 cells and its mechanism. Methods Cell counting reagent (CCK-8) was used to detect the effect of Sch B on the survival rate of MDA-MB-231 cells. MDA-MB-231 cells were treated with Sch B (10, 20, 40 μmol/L) for 24 hours. The cell death was detected by Annexin V-FITC/PI. The levels of intracellular reactive oxygen species (ROS) were detected by DCFA-DA fluorescent probe. Apoptosis and the expression of endoplasmic reticulum stress related proteins (Bcl-2、Bax、CHOP、GPR78、PERK、p-PERK、p-eIF2α、eIF2) were detected by Western blot. Results Compared with the blank group, the cell survival rate decreased significantly (P<0.01) with the increase of Sch B concentration, and its IC50 was 19.16 μmol/L. Compared with the control group, Sch B groups (10, 20, 40 μmol/L) inhibited cell clone formation in a dose-dependent manner (P<0.05). Sch B groups (10, 20, 40 μmol/L) induced apoptosis (P<0.05), significantly reduced the expression of anti-apoptotic protein Bcl-2 and significantly increased the expression of pro-apoptotic protein Bax (P<0.05). Sch B groups (10, 20, 40 μmol/L) significantly increased the level of intracellular ROS in a dose-dependent manner (P<0.05). Sch B groups (10, 20, 40 μmol/L) stimulated endoplasmic reticulum stress and increased the expressions of endoplasmic reticulum stress-related proteins CHOP, GPR78 and p-eIF2α in a dose-dependent manner (P<0.05). Conclusion Sch B induces apoptosis of MDA-MB-231 cells through ROS mediated endoplasmic reticulum stress. -
Key words:
- schisandrin B /
- breast cancer /
- reactive oxygen species /
- endoplasmic reticulum stress /
- apoptosis
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