人参皂苷S-Rh2诱导卵巢癌SKOV3细胞自噬作用及机制的实验研究

发布时间：2018-05-20 11:17

本文选题：人参皂苷S-Rh2 + 卵巢癌　；参考：《吉林大学》2017年硕士论文

【摘要】：目的:研究S-Rh2诱导的SKOV3细胞株自噬变化,初步探索PI3K/AKT/m TOR通路在本实验中起到的调控作用,并进一步探讨其调控机制。方法:1.CCK8法测定S-Rh2单独应用及以PI3K/m TOR阻断剂预处理后对SKOV3细胞的增殖抑制作用。2.观察不同药物处理条件下SKOV3细胞形态变化,吖啶橙染色后荧光显微镜下检测其自噬溶酶体形成情况。3.Western-blot法检测S-Rh2处理后PI3K/AKT/m TOR通路蛋白表达与SKOV3细胞自噬之间的关联。结果:1.当浓度在40-80μg/ml范围内时,人参皂苷S-Rh2对人卵巢癌SKOV3细胞具有增殖抑制作用,并呈剂量依赖性。在加入1μg/ml NVP-BEZ235后,S-Rh2浓度为10μg/ml、20μg/ml、40μg/ml组的增殖抑制作用均显著提高,P0.01,浓度为80ug/ml组则P0.05无统计学意义。2.S-Rh2作用后的SKOV3细胞P-AKT、AKT、m TOR蛋白的表达下降,出现PI3K/AKT/m TOR通路的抑制,该现象与S-Rh2的浓度呈正相关,并与自噬蛋白LC3 II/LC3 I呈正相关。3.吖啶橙染色可见随S-Rh2浓度增高,SKOV3细胞内自噬溶酶体逐渐增加,贴壁细胞密度降低,细胞质皱缩变圆,而阻断PI3K/AKT/m TOR通路后,SKOV3细胞内自噬溶酶体红色荧光强度高于阻断前。结论:人参皂苷S-Rh2在杀伤卵巢癌SKOV3细胞的同时能够诱导其PI3K/AKT/m TOR通路抑制并产生自噬,该变化与S-Rh2的作用浓度呈现正相关。在应用NVP-BEZ235阻断PI3K/AKT/m TOR通路后,S-Rh2对SKOV3细胞的增殖抑制作用在一定程度上有提高,同时伴随着相应细胞的自噬程度增加。
[Abstract]:Aim: to study the changes of autophagy induced by S-Rh2 in SKOV3 cell lines, and to explore the regulatory role of PI3K/AKT/m TOR pathway in this experiment and its regulatory mechanism. Methods 1. CCK8 assay was used to determine the inhibitory effect of S-Rh2 on the proliferation of SKOV3 cells after being used alone and pretreated with PI3K/m TOR blocker. Morphological changes of SKOV3 cells were observed under different drug treatment conditions. Lysosome formation of SKOV3 cells was detected by fluorescence microscope after acridine orange staining. 3. Western-blot method was used to detect the relationship between PI3K/AKT/m TOR pathway protein expression and autophagy of SKOV3 cells after S-Rh2 treatment. The result is 1: 1. Ginsenoside S-Rh2 inhibited the proliferation of human ovarian cancer SKOV3 cells in a dose-dependent manner when the concentration was in the range of 40-80 渭 g/ml. After adding 1 渭 g/ml NVP-BEZ235, the proliferation inhibition of 10 渭 g / ml ~ 20 渭 g 路ml ~ (-1) L ~ (20 渭 g 路L ~ (-1) 路L ~ (-1) g/ml) was significantly higher than that of 80ug/ml group (P 0.05). 2.The expression of P-AKTK ~ (TM) TOR in SKOV3 cells treated with S-Rh2 was decreased, and the PI3K/AKT/m TOR pathway was inhibited. This phenomenon was positively correlated with the concentration of S-Rh2 and with the autophagy protein LC3 II/LC3 I. Acridine orange staining showed that with the increase of S-Rh2 concentration, the lysosome in SKOV3 cells increased gradually, the density of adherent cells decreased, the cytoplasm became round, and the red fluorescence intensity of autophagy lysosome in SKOV3 cells was higher than that before blocking PI3K/AKT/m TOR pathway. Conclusion: ginsenoside S-Rh2 can induce inhibition of PI3K/AKT/m TOR pathway and autophagy in ovarian cancer SKOV3 cells, which is positively correlated with the concentration of S-Rh2. After blocking the PI3K/AKT/m TOR pathway by NVP-BEZ235, the inhibitory effect of S-Rh2 on the proliferation of SKOV3 cells was increased to a certain extent, and the autophagy degree of the corresponding cells was increased.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.31

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