【作者】 陈宇；

【导师】 蔡红兵；

【作者基本信息】 武汉大学 ， 妇产科学， 2022， 硕士

【摘要】 第一部分氯喹对卵巢癌细胞增殖、迁移和自噬的影响目的:探讨氯喹(CQ)对卵巢癌A2780细胞和SKOV3细胞增殖、迁移和自噬的影响,并对其产生作用的机制进行研究。方法:1、采用CCK8法检测不同浓度氯喹处理A2780细胞和SKOV3细胞后的增殖能力;2、克隆形成实验检测氯喹对A2780细胞和SKOV3细胞克隆形成能力的影响;3、划痕实验和Transwell实验检测氯喹对A2780细胞和SKOV3细胞的迁移能力的影响;4、红绿双荧光m RFP-e GFP-LC3自噬双标质粒转染A2780细胞和SKOV3细胞,荧光显微镜观察氯喹处理48h后自噬小体的形成和自噬流的变化;5、Western blot检测氯喹处理A2780细胞和SKOV3细胞48h后自噬标志蛋白LC3的表达及PI3K/Akt/m TOR通路相关蛋白的表达。结果:1、氯喹显著抑制A2780细胞和SKOV3细胞增殖能力和迁移能力,氯喹处理能显著抑制卵巢癌细胞的自噬能力。2、A2780细胞和SKOV3细胞经氯喹处理后LC3Ⅱ、PI3K、p-AKT和m TOR蛋白表达水平显著增高,而AKT、p-m TOR蛋白水平显著降低。结论:氯喹能显著抑制A2780细胞和SKOV3细胞增殖、迁移能力和自噬,抑制细胞自噬的机制可能是通过激活PI3K/Akt/m TOR通路。第二部分氯喹对卵巢癌细胞化疗敏感性影响目的:初步探讨氯喹(CQ)对卵巢癌A2780细胞和SKOV3细胞的顺铂增敏作用。方法:1、采用CCK8法检测氯喹联合顺铂治疗对A2780细胞和SKOV3细胞后增殖能力的影响;2、克隆形成实验检测氯喹联合顺铂治疗对A2780细胞和SKOV3细胞克隆形成的影响;3、划痕实验和Transwell实验检测氯喹联合顺铂治疗对A2780细胞和SKOV3细胞的迁移能力的影响。结果:1、相比单独使用顺铂,顺铂联合氯喹显著抑制A2780细胞和SKOV3细胞增殖能力。2、相比单独使用顺铂,顺铂联合氯喹显著抑制A2780细胞和SKOV3细胞迁移能力。结论:氯喹能显著增强卵巢癌细胞对顺铂的化疗敏感性。更多还原

【Abstract】 Part Ⅰ Effects of chloroquine on proliferation,migration and autophagy of ovarian cancer cellsObjective To investigate the effect of chloroquine(CQ)on the proliferation,migration and autophagy of ovarian cancer A2780 cells and SKOV3 cells,and to study the mechanism of its effect.Methods 1.The proliferation ability of A2780 cells and SKOV3 cells treated with different concentrations of chloroquine was detected by CCK8 method.2.The clone formation assay was used to detect the effect of chloroquine(15 μmol/L)on the colony formation of A2780 cells and SKOV3 cells.3.The scratch assay and Transwell assay were used to detect the migration ability of A2780 cells and SKOV3 cells treated with chloroquine(15 μmol/L).4.Red and green double fluorescence m RFP-e GFP-LC3 autophagy double-labeled plasmids were transfected into A2780 cells and SKOV3 cells,and fluo-rescence microscope was adopted to observe the formation of autophagosomes and changes in autophagic flux after chloroquine(15 μmol/L)treatment for 48 h.5.Western blot was used to detect the expression of autophagy marker protein LC3 and the expression of PI3K/Akt/m TOR pathway-related proteins in A2780 cells and SKOV3 cells treated with chloroquine for 48 h.Results 1.Chloroquine significantly inhibited the proliferation and migration of A2780 cells and SKOV3 cells.In addition,chloroquine treatment could significantly inhibit the autophagy of ovarian cancer cells.2.After A2780 cells and SKOV3 cells were treated with chloroquine,the protein expressions of LC3Ⅱ,PI3 K,p-AKT and m TOR were significantly increased,while the protein levels of AKT and p-m TOR were significantly decreased..Conclusion Chloroquine can significantly inhibit the proliferation and migration of A2780 cells and SKOV3 cells,and may inhibit autophagy by activating the PI3K/Akt/m TOR pathway.Part Ⅱ Chloroquine affects the chemosensitivity of ovarian cancer cells to cisplatin Objective To preliminarily investigate the cisplatin-sensitizing effect of chloroquine(CQ)on ovarian cancer A2780 cells and SKOV3 cells.Methods 1.The proliferation ability of A2780 cells and SKOV3 cells treated with cisplatin with or without chloroquine was detected by CCK8 method;2.The clone formation assay was used to detect the effect of cisplatin with or without chloroquine on the clone formation of A2780 cells and SKOV3 cells;3.The effect of cisplatin with or without chloroquine on the migration ability of A2780 cells and SKOV3 cells was detected by scratch test and Ttranswell test.Results: 1.Compared with cisplatin alone,cisplatin combined with chloroquine significantly inhibited the proliferation of A2780 cells and SKOV3 cells.2.Compared with cisplatin alone,cisplatin combined with chloroquine significantly inhibited the migration ability of A2780 cells and SKOV3 cells.Conclusion: Chloroquine can significantly affect the chemosensitivity of ovarian cancer cells to cisplatin.更多还原