BACKGROUND Cordyceps militaris (L.) Fr. (C. militaris) exhibits pharmacological activities, including antitumor properties, through the regulation of the nuclear factor kappa B (NF-κB) signaling. Tumor Necrosis Factor (TNF) and TNF-α modulates cell survival and apoptosis through NF- κB signaling. However, the mechanism underlying its mode of action on the NF-κB pathway is unclear. METHODS Here, we analyzed the effect of C. militaris extract (CME) on the proliferation of ovarian cancer cells by confirming viability, morphological changes, migration assay. Additionally, CME induced apoptosis was determined by apoptosis assay and apoptotic body formation under TEM. The mechanisms of CME were determined through microarray, immunoblotting and immunocytochemistry. RESULTS CME reduced the viability of cells in a dose-dependent manner and induced morphological changes. We confirmed the decrease in the migration activity of SKOV-3 cells after treatment with CME and the consequent induction of apoptosis. Immunoblotting results showed that the CME-mediated upregulation of tumor necrosis factor receptor 1 (TNFR1) expression induced apoptosis of SKOV-3 cells via the serial activation of caspases. Moreover, CME negatively modulated NF-κB activation via TNFR expression, suggestive of the activation of the extrinsic apoptotic pathway. The binding of TNF-α to TNFR results in the disassociation of IκB from NF-κB and the subsequent translocation of the active NF-κB to the nucleus. CME clearly suppressed NF-κB translocation induced by interleukin (IL-1β) from the cytosol into the nucleus. The decrease in the expression levels of B cell lymphoma (Bcl)-xL and Bcl-2 led to a marked increase in cell apoptosis. CONCLUSION These results suggest that C. militaris inhibited ovarian cancer cell proliferation, survival, and migration, possibly through the coordination between TNF-α/TNFR1 signaling and NF-κB activation. Taken together, our findings provide a new insight into a novel treatment strategy for ovarian cancer using C. militaris.

中文翻译：

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虫草通过TNF-α/ TNFR1介导的NF-κB磷酸化抑制作用，诱导卵巢癌细胞凋亡。

背景Cord虫草（L.）Fr. C. militaris（C. militaris）通过调节核因子κB（NF-κB）信号传导表现出药理活性，包括抗肿瘤特性。肿瘤坏死因子（TNF）和TNF-α通过NF-κB信号传导调节细胞存活和凋亡。但是，尚不清楚其作用于NF-κB途径的机制。方法在这里，我们通过确认生存力，形态变化和迁移分析，分析了C.militaris提取物（CME）对卵巢癌细胞增殖的影响。另外，通过在TEM下的细胞凋亡测定和凋亡小体形成来确定CME诱导的细胞凋亡。通过微阵列，免疫印迹和免疫细胞化学确定了CME的机制。结果CME以剂量依赖性方式降低了细胞的活力并诱导了形态学改变。我们证实了用CME处理后SKOV-3细胞迁移活性的降低以及随后的凋亡诱导。免疫印迹结果表明，CME介导的肿瘤坏死因子受体1（TNFR1）表达的上调通过胱天蛋白酶的系列激活诱导SKOV-3细胞凋亡。此外，CME通过TNFR表达负调节NF-κB的激活，提示外在凋亡途径的激活。TNF-α与TNFR的结合导致IκB与NF-κB分离，并随后将活性NF-κB转运至细胞核。CME明显抑制了白介素（IL-1β）诱导的NF-κB从胞浆进入细胞核。B细胞淋巴瘤（Bcl）-xL和Bcl-2的表达水平降低导致细胞凋亡明显增加。结论这些结果表明，游衣藻可能通过TNF-α/ TNFR1信号传导与NF-κB活化之间的协同作用来抑制卵巢癌细胞的增殖，存活和迁移。综上所述，我们的发现为使用C.militaris的卵巢癌新型治疗策略提供了新见解。