【作者】 徐蕾；

【导师】 钟玲；

【作者基本信息】 重庆医科大学 ， 妇产科学， 2010， 硕士

【摘要】 目的观察在不同浓度葡萄籽原花青素的干预下,人卵巢癌移植瘤小鼠皮下移植瘤的生长情况及其肿瘤细胞在形态学上的变化,检测葡萄籽原花青素对肿瘤细胞凋亡情况的影响,探讨其对肿瘤细胞中各凋亡基因在分子和蛋白水平表达的改变,分析葡萄籽原花青素影响人卵巢癌移植瘤小鼠肿瘤细胞的增殖和凋亡的可能机制。方法(1)体外培养人卵巢癌SKOV3细胞,接种至24只裸鼠皮下,建立人卵巢癌裸鼠皮下移植瘤模型,随机分成3组,每组8只:原花青素高浓度组(200 mg/kg),原花青素低浓度组(100 mg/kg)及空白对照组。裸鼠每天以原花青素溶液或生理盐水灌胃,定期观察各组裸鼠移植瘤生长情况,测量其肿瘤体积。(2)处理结束后处死裸鼠,取肿瘤组织进行石蜡切片HE染色观察肿瘤细胞形态,流式细胞分析细胞周期及凋亡。(3)RT-PCR,Westrnblot检测肿瘤细胞的Caspase-3,Bax,Bcl-2基因在RNA及蛋白水平的表达情况。结果(1)原花青素处理后的肿瘤较对照组生长缓慢,瘤体较小,瘤重较轻,原花青素高、低浓度组皮下移植瘤的抑瘤率及瘤重与对照组相比差异具有显著性。(2)肿瘤组织进行石蜡切片HE染色观察,肿瘤细胞出现明显的凋亡形态学变化。流式细胞检测各组肿瘤细胞的周期分布显示:经原花青素处理后,肿瘤细胞被阻滞在G2/M期,Annexin-V和PI染色后FCM分析细胞凋亡表明,原花青素处理后的肿瘤组织出现了较多凋亡和死亡的细胞,原花青素高、低浓度组的细胞凋亡率较对照组差异有显著性。(3)原花青素可激活Caspase-3的表达活性,明显增高Bax,降低Bcl-2基因在RNA和蛋白水平的表达,且其改变与原花青素的处理浓度相关。结论(1)葡萄籽原花青素对人卵巢癌裸鼠皮下移植瘤的生长有抑制作用。(2)葡萄籽原花青素对移植瘤肿瘤细胞的凋亡有促进作用。(3)葡萄籽原花青素可能通过增强Bax,减低Bcl-2的表达,激活Caspase-3途径而发挥效应。更多还原

【Abstract】 Objective To observe the subcutaneous tumor growth and tumor cell morphological changes of human ovarian cancer in nude mice at different concentrations of Grape seed proanthocyanidin. Detect the changes on tumor cell apoptosis and the apoptosis gene expression at the molecular and protein level, and to explore the molecular mechanism on proliferation and apoptosis of the human ovary cancer in nude mice.Methods (1)A human ovarian cancer model subcutaneously transplanted into nude mice was established.They were divided into 3 groups:the HD GSP group,the LD GSP group and the control group,8 in each.Tumor volume and weight were detected. (2)HE stain was used to observe the tumor cells morphous. Apoptosis of the tumor cells was analyzed by flowcytometry (FCM) (3), semi - quantitative RT-PCR was used to detect the mRNA expression of Caspase-3,Bax,Bcl-2 and the protein expression of them were detected by Westernblot.Results (1) The GSP treated tumors grew slower and smaller than the control group one,the inhibitory rate in the HD and LD GSP groups were significantly higher than that in the control group. (2) Tumor cells show obvious morphological changes of apoptosis by HE stained paraffin sections. Flow cytometry to detect cell cycle distribution of tumor found that the tumor cells were arrested in G2 / M phase, and the apoptotic rate in the GSP groups were significantly higher than that in the control group by Annexin-V and PI staining FCM analysis . (3)Semiquantitative RT-PCR and Western blot showed the Caspase-3, Bax mRNA expression and protein expression were both up-regulated,and the Bcl-2 expression was in reverse.Conclusion (1)GSP can inhibit the growth of ovarian cancer in nude mice.(2)GSP can induce the apoptosis of tumor cells.(3)GSP may induce the cell apoptosis by means of up-regulating the expression of Bax,Caspase-3 and decreasing the expression of Bcl-2更多还原