Department of Obstetrics and Gynecology, the First Affiliated Hospital, Harbin Medical University, Harbin 150001, China.
Chin J Cancer Res. 2011 Jun;23(2):140-6. doi: 10.1007/s11670-011-0140-1.
The Notch signaling pathway plays an important role in the stem cell signaling network and contributes to tumorigenesis. However, the functions of Notch signaling in ovarian cancer stem cells (OCSCs) are not well understood. We aimed to investigate the effects of Notch blockade on self-renewal and stemness maintenance of OCSCs.
Ovarian cancer stem-like cells were enriched from ovarian cancer cell lines in serum-free medium. A γ-secretase inhibitor, (DAPT), was used to block Notch signaling. MTT assays were performed to assess self-renewal and proliferation inhibition, flow cytometry was performed to analyze cell surface marker and immunofluorescence, Western Blot and Real-time RT-PCR assays were performed to detect Oct4 and Sox2 protein and mRNA expression of the Ovarian cancer stem-like cells treated with DAPT.
Notch blockade markedly inhibits self-renewal and proliferation of ovarian cancer stem-like cells, significantly downregulates the expression of OCSCs-specific surface markers, and reduces protein and mRNA expression of Oct4 and Sox2 in OCSC-like cells.
Our results suggest that Notch signaling is not only critical for the self-renewal and proliferation of OCSCs, but also for the stemness maintenance of OCSCs. The γ-secretase inhibitor is a promising treatment targeting OCSCs.
Notch 信号通路在干细胞信号网络中发挥着重要作用，并有助于肿瘤的发生。然而，Notch 信号在卵巢癌干细胞（OCSCs）中的功能还不是很清楚。我们旨在研究 Notch 阻断对 OCSCs 自我更新和干性维持的影响。
在无血清培养基中从卵巢癌细胞系中富集卵巢癌干细胞样细胞。使用γ-分泌酶抑制剂（DAPT）阻断 Notch 信号。通过 MTT 检测评估自我更新和增殖抑制，通过流式细胞术分析细胞表面标志物，通过免疫荧光、Western Blot 和 Real-time RT-PCR 检测 DAPT 处理后的 OCSC 样细胞中 Oct4 和 Sox2 蛋白和 mRNA 的表达。
Notch 阻断显著抑制卵巢癌干细胞样细胞的自我更新和增殖，显著下调 OCSCs 特异性表面标志物的表达，并降低 OCSC 样细胞中 Oct4 和 Sox2 的蛋白和 mRNA 表达。
我们的结果表明，Notch 信号不仅对 OCSCs 的自我更新和增殖至关重要，而且对 OCSCs 的干性维持也至关重要。γ-分泌酶抑制剂是一种有前途的针对 OCSCs 的治疗靶点。
