Department of Obstetrics and Gynecology, CHA Gangnam Medical Center, CHA University School of Medicine, Seoul, Republic of Korea.
Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital, Seongnam, Republic of Korea.
Biomed Pharmacother. 2018 Dec;108:584-589. doi: 10.1016/j.biopha.2018.09.041. Epub 2018 Sep 20.
Bmi1, a polycomb group gene, is essential for self-renewal of stem cells and is frequently upregulated in various cancer cells. We aimed to investigate the effect of Bmi1 silencing on cancer stemness and chemosensitivity in endometrial cancer using targeted siRNA approach in HEC1A and Ishikawa cells.
Cell viability after treatment with Bmi1 siRNA was assessed using the MTT assay, and cell apoptosis was visualized using the TdT-mediated dUTP nick-end labeling (TUNEL) method. Western blotting, migration assays and invasion assays were performed to detect changes in the stem-like properties of cancer cells. To evaluate the anticancer effect of Bmi1 silencing, HEC1A and Ishikawa cells were treated with 100 nM Bmi1 siRNA and/or 40 μM cisplatin.
In the MTT assay, compared to control, viability of HEC1A and Ishikawa cells significantly decreased after Bmi1 siRNA treatment in a dose-dependent manner. Bmi1 silencing using siRNA increased the expression of cleaved caspase-3 and cleaved poly adenosine diphosphate-ribose polymerase polymerase (PARP) as observed in the western blot analysis. Apoptosis significantly increased in the HEC1A and Ishikawa cells treated with 100 nM Bmi1 siRNA for 48 h than in the control cells in TUNEL assay. SOX2 and Oct4 expression decreased in the HEC1A and Ishikawa cells treated with Bmi1 siRNA, while E-cadherin expression increased. Further, migratory and invasive properties were significantly inhibited by Bmi1 siRNA treatment in both cell lines. Notably, viability of HEC1A and Ishikawa cells decreased more when they were concurrently treated with Bmi1 siRNA and cisplatin compared to when they were treated with Bmi1 siRNA or cisplatin alone.
Bmi1 silencing suppresses cancer stemness in HEC1A and Ishikawa cells. Concurrent treatment with Bmi1 siRNA and cisplatin resulted in additive anticancer effect with a cell line-specific pattern, which was higher than that shown by cisplatin treatment alone.
BMI1 是多梳组基因家族的一员，对于干细胞的自我更新至关重要，并且在多种癌细胞中经常上调。我们旨在使用靶向 siRNA 方法在 HEC1A 和 Ishikawa 细胞中研究 BMI1 沉默对子宫内膜癌细胞干性和化疗敏感性的影响。
使用 MTT 测定法评估用 BMI1 siRNA 处理后的细胞活力，并用 TdT 介导的 dUTP 缺口末端标记法（TUNEL）可视化细胞凋亡。进行 Western blot、迁移和侵袭试验以检测癌细胞干性样特性的变化。为了评估 BMI1 沉默的抗癌作用，用 100 nM BMI1 siRNA 和/或 40 μM 顺铂处理 HEC1A 和 Ishikawa 细胞。
在 MTT 测定中，与对照组相比，HEC1A 和 Ishikawa 细胞在用 siRNA 处理后，细胞活力呈剂量依赖性显著降低。Western blot 分析显示，siRNA 介导的 BMI1 沉默增加了 cleaved caspase-3 和 cleaved poly adenosine diphosphate-ribose polymerase 聚合酶（PARP）的表达。在用 100 nM BMI1 siRNA 处理 48 小时后，TUNEL 测定中凋亡在 HEC1A 和 Ishikawa 细胞中显著增加。与对照组相比，SOX2 和 Oct4 在接受 BMI1 siRNA 处理的 HEC1A 和 Ishikawa 细胞中的表达降低，而 E-cadherin 的表达增加。此外，在两种细胞系中，BMI1 siRNA 处理均显著抑制迁移和侵袭特性。值得注意的是，与单独用 BMI1 siRNA 或顺铂处理相比，当同时用 BMI1 siRNA 和顺铂处理时，HEC1A 和 Ishikawa 细胞的活力下降更多。
BMI1 沉默抑制了 HEC1A 和 Ishikawa 细胞的癌症干性。与单独用顺铂处理相比，BMI1 siRNA 与顺铂的联合治疗具有细胞系特异性的附加抗癌作用，其作用高于单独用顺铂处理。
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