Berberine in combination with cisplatin induces necroptosis and apoptosis in ovarian cancer cells
Berberine in combination with cisplatin induces necroptosis and apoptosis in ovarian cancer cells
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DOI：
10.1186/s40659-019-0243-6
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发表时间：
2019-07-18
影响因子：
6.7
通讯作者：
Cheng, Zhongping
中科院分区：
文献类型：
--
作者：
Liu, Li;Fan, Jingyan;Cheng, Zhongping
背景资料：小檗碱（BBR）是一种从多种中草药中提取的化合物，对不同来源的癌细胞具有多种药理作用。顺铂（DDP）是一种有效的抗癌化疗药物，其作用机制是诱导DNA损伤和细胞凋亡。方法：以OVCAR 3和3株患者来源的卵巢癌原代细胞系（POCCLs）为实验对象，采用流式细胞术（FCM）、流式细胞术（FCM）等方法，观察BBR联合DDP对卵巢癌细胞凋亡的影响。为了确定BBR和DDP组合的潜在抗癌活性，我们首先用BBR和/或DDP处理OVCAR 3和POCCL细胞。通过CCK-8测定法测量BBR或DDP处理不同时间的OVCAR 3和POCCL的细胞活力。流式细胞仪检测BBR和/或DDP处理后细胞周期分布及凋亡细胞的变化。透射电镜观察OVCAR 3细胞形态学变化。RT-qPCR、Western blotting和免疫荧光法检测BBR或DDP处理OVCAR 3和POCCLs后细胞增殖、凋亡和坏死相关标志物的变化。BBR与DDP联合应用对肿瘤细胞生长有明显的抑制作用，并可诱导肿瘤细胞阻滞于G 0/G1期。透射电镜观察发现，BBR或DDP处理后的细胞大多数呈典型的凋亡和坏死细胞死亡形态。结论：BBR和DDP联合应用可诱导卵巢癌细胞凋亡和坏死，从而提高化疗药物的抗肿瘤效果。凋亡涉及caspase依赖性途径，而坏死性凋亡涉及RIPK 3-MLKL途径的激活。我们希望我们的研究结果可以为BBR作为卵巢癌治疗药物的潜力提供新的见解。
Background: Berberine (BBR), a compound extracted from a variety of medicinal herbs, has been shown multiple pharmacological effects against cancer cells of different origins. Cisplatin (DDP) is known as an effective chemotherapeutic agent against cancerby inducing DNA damage and cell apoptosis. However, the effect of the combined used of BBR and DDP on cell necroptosis in ovarian cancer has not been reported.Methods: OVCAR3 and three patient-derived primary ovarian cancer cell lines (POCCLs) were chosen as the experimental objects. To determine the potential anti-cancer activity of BBR and DDP in combination, we firstly treated OVCAR3 and POCCLs cells with BBR and/or DDP. The cell viability of OVCAR3 and POCCLs with treatment of BBR or DDP for different hours was measured by CCK-8 assay. Flow cytometry was used to analyze cell cycle distribution and changes in apoptotic cells after treatment with BBR and/or DDP. The morphological changes of OVCAR3 cells were observed by using Transmission electron microscopy (TEM) analysis. Proliferation, apoptosis and necroptosis related markers of OVCAR3 and POCCLs with treatment of BBR or DDP were measured by RT-qPCR, western blotting and immunofluorescence assay.Results: Our results demonstrated that BBR significantly inhibited the proliferation of OVCAR3 and primary ovarian cancer cells in a dose- and time-dependent manner. The combination treatment of BBR and DDP had a prominent inhibitory effect on cancer cell growth and induced G0/G1 cell cycle arrest. TEM revealed that the majority of cells after BBR or DDP treatment had an increasing tendency of typical apoptotic and necrotic cell death morphology. Besides, BBR and DDP inhibited the expression of PCNA and Ki67 and enhanced the expression and activation of Caspase-3, Caspase-8, RIPK3 and MLKL.Conclusion: This study proposed that the combination therapy of BBR and DDP markedly enhanced more ovarian cancer cell death by inducing apoptosis and necroptosis, which may improve the anticancer effect of chemotherapy drugs. The apoptosis involved the caspase-dependent pathway, while the necroptosis involved the activation of the RIPK3-MLKL pathway. We hope our findings might provide a new insight for the potential of BBR as a therapeutic agent in the treatment of ovarian cancer.