Jin Zhishan, Zheng Lang, Xin Xiaoyan, Li Yuanyue, Hua Teng, Wu Tingting, Wang Hongbo
Department of Obstetrics and Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, P.R. China.
Department of Obstetrics and Gynecology, Hainan Provincial People's Hospital, Haikou, Hainan, P.R. China.
Mol Med Rep. 2014 Dec;10(6):3099-105. doi: 10.3892/mmr.2014.2664. Epub 2014 Oct 15.
Ceramide is a bioactive lipid which functions as a tumor suppressor, mediating processes such as apoptosis, growth arrest, senescence and differentiation. The effects of ceramide in ovarian cancers have not been well established. The objective of the present study was to investigate the effects of C2‑ceramide treatment in A2780 ovarian cancer cells and its possible molecular mechanism. C2‑ceramide-induced proliferation inhibition was analyzed using an MTT assay and Trypan blue test. Flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling were used to identify the induction of apoptosis. Transmission electron microscopy was used to confirm the formation of autophagosomes. Quantitative polymerase chain reaction was performed to analyze the messenger RNA expression of the autophagy and cell death associated genes and western blotting was used to analyze the protein expression of beclin 1, LC3, Akt, forkhead box O3 (FOXO3) and adenosine monophosphate-activated protein kinase in ovarian cancer cells. It was found that C2‑ceramide inhibited A2780 cell proliferation in a time‑ and dose‑dependent manner and C2‑ceremide induced A2780 cell apoptosis and autophagy. However, C2‑ceramide‑induced autophagy did not result in cell death, but instead protected ovarian cancer cells from apoptosis. Akt inhibition and FOXO3 activation were implicated in C2‑ceramide‑treated ovarian cancer cells. Furthermore, FOXO3 target genes, which were associated with autophagy (MAP1LC3, GABARAP and GABARAPL1) and cell death (BNIP3, BNIP3L, BIM and PUMA), were upregulated. The present study has shown that C2‑ceramide induced apoptosis and autophagy in ovarian cancer cells. FOXO3 transcription was upregulated, which may contribute to C2‑ceramide‑induced apoptosis and autophagy.
神经酰胺是一种生物活性脂质，具有肿瘤抑制功能，介导细胞凋亡、生长停滞、衰老和分化等过程。神经酰胺在卵巢癌中的作用尚未完全明确。本研究的目的是探讨C2-神经酰胺处理对A2780卵巢癌细胞的影响及其可能的分子机制。采用MTT法和台盼蓝试验分析C2-神经酰胺诱导的增殖抑制作用。通过流式细胞术和末端脱氧核苷酸转移酶dUTP缺口末端标记法鉴定细胞凋亡的诱导情况。利用透射电子显微镜确认自噬体的形成。进行定量聚合酶链反应分析自噬和细胞死亡相关基因的信使核糖核酸表达，并采用蛋白质印迹法分析卵巢癌细胞中Beclin 1、LC3、Akt、叉头框O3（FOXO3）和腺苷酸活化蛋白激酶的蛋白表达。结果发现，C2-神经酰胺以时间和剂量依赖性方式抑制A2780细胞增殖，并诱导A2780细胞凋亡和自噬。然而，C2-神经酰胺诱导的自噬并未导致细胞死亡，反而保护卵巢癌细胞免受凋亡。Akt抑制和FOXO3激活与C2-神经酰胺处理的卵巢癌细胞有关。此外，与自噬（MAP1LC3、GABARAP和GABARAPL1）和细胞死亡（BNIP3、BNIP3L、BIM和PUMA）相关的FOXO3靶基因上调。本研究表明，C2-神经酰胺诱导卵巢癌细胞凋亡和自噬。FOXO3转录上调，这可能有助于C2-神经酰胺诱导的凋亡和自噬。
