神经酰胺通过akt1foxo3a信号通路诱导卵巢癌细胞自噬及其机制的分析-ceramide induces autophagy of ovarian cancer cells through ak t1 foxo 3a signal pathway and its mechanism analysis.docxVIP

华中科技大学硕士学位论文 华 中 科 技 大 学 硕 士 学 位 论 文 5 5 C2-ceramide elevated, the IC50 value was 55.719μmol/L; The rate of early apoptosis of the ovarian cancer cells depended on the concentrations of C2-ceramide. But, for the concentration of C2-ceramide less than 25μmol/L, there was almost no initiation of ovarian cancer cells apoptosis. Transmission electron microscopy showed autophagic vacuoles of A2780 cells treated with C2-ceramide. With the supplement of C2-ceramide, the mRNA and protein expression of Beclin1 and LC3 in the ovarian cancer cells were increased. LC3 had two subunits, LC3 I and LC3 II .The expressions of LC3 I and LC3 II protein were increased. However , there was a greater increase in LC3II than LC3I. P-Akt1 (ser473) protein expression level decreased with the addition of C2-ceramide, meanwhile the Fox3a protein expression level was increased. Conclusions: C2-ceramide could induce ovarian cancer cells autophagic death through Akt1 /Foxo3a signaling pathway. C2-ceramide could activate autophagy- related genes Beclin1 and LC3 expression. 【Keyword】C2-ceramide; LC3; Beclin1; autophay; Akt1; Foxo3a 1 1 英文缩略词表 Cer Akt1 Foxo3a WB MDR PBS PCR Q-PCR Beclin1 LC3 ceramide protein kinaseB1 forkhead box3a western blot multiple drug resistance phosphate buffer saline polymerase chain reaction Real time quantitative PCR Beclin1 microtubule-associated protein1 light chain3 神经酰胺 磷酸化蛋白激酶 B1 叉头框蛋白 3a 蛋白免疫印迹 多重耐药 磷酸盐缓冲液 聚合酶链式反应 荧光定量聚合酶链式反应 自噬相关蛋白 微管相关蛋白 1 轻链 3 dNTP cDNA FCM PCD ATG PE TEM deoxyribonucleotide triphosphate complementary deoxyribonucleic acid Flow cycometer Programmed cell death autophagy-related gene phosphatidylethanolamine transmission electron microscropy 脱氧核糖核苷三磷酸 互补脱氧核糖核酸 流式细胞仪 程序性细胞死亡 自噬相关基因 磷脂酰乙醇胺 透射电子显微镜 独创性声明 本人郑重声明，本学位论文是本人在导师指导下进行的研究工作及取得的研究成 果的总结。尽我所知，除文中已经标明引用的内容外，本论文不包含任何其他个人或 集体已经发表或撰写过的研究成果。对本文的研究做出贡献的个人和集体，均已在文 中以明确方式标明。本人完全意识到本人将承担本声明引起的一切法律后果。 学位论文作者签名： 日期： 年 月 日 学位论文版权使用授权书 本学位论文作者完全了解学校有关保留、使用学位论文的规定，即：学校有权保 留并向国家有关部门或机构送交论文的复印件和电子版，允许论文被查阅和借阅