两种抗癌中药与顺铂联用对人乳腺癌MCF-7细胞系增殖抑制作用及机制的研究

发布时间：2018-03-13 03:31

本文选题：去甲斑蝥酸钠　切入点：顺铂　出处：《吉林大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:本研究通过研究不同浓度的传统中药姜黄素、去甲斑蝥酸钠和化疗药物顺铂在不同时间,单药以及联合用药对人乳腺癌MCF-7细胞增殖抑制的作用以及诱导凋亡和细胞周期的影响,初步探讨去甲斑蝥酸钠与顺铂联用抑制人乳腺癌MCF-7细胞株可能的作用机制,同时对比单独用药与联合用药细胞增殖抑制作用的情况,从而为乳腺癌治疗策略提供有效的基础实验数据做参考。方法:1、人乳腺癌细胞MCF-7细胞体外培养至对数生长期。2、通过MTT法检测不同浓度的姜黄素、去甲斑蝥酸钠和顺铂以及两药联合作用于MCF-7细胞24、48和72小时后细胞增殖的情况。3、使用流式细胞仪检测去甲斑蝥酸钠(30μg/ml、15μg/ml)、顺铂(1.25μg/ml、0.625μg/ml)单独以及联合作用于MCF-7细胞48小时后诱导凋亡的情况和细胞周期的影响。结果:1.姜黄素、去甲斑蝥酸钠和顺铂对MCF-7细胞增殖的抑制情况去甲斑蝥酸钠和顺铂对MCF-7细胞均有增殖抑制作用,并且抑制效果呈时间-浓度依赖性的表现,姜黄素通过实验的统计结果来看对MCF-7无明显抑制作用。2.不同浓度去甲斑蝥酸钠与顺铂联用对人乳腺腺癌MCF-7细胞增殖的抑制作用联合应用去甲斑蝥酸钠(30μg/ml、15μg/ml)和顺铂(1.25μg/ml、0.625μg/ml),对人乳腺癌MCF-7细胞的抑制作用均高于单独用药组,随着去甲斑蝥酸钠和顺铂的浓度增加,抑制作用增强,同时,单纯的增加去甲斑蝥酸钠的浓度,对MCF-7的抑制作用也在增强,随着作用时间的延长抑制作用增强。3.去甲斑蝥酸钠、顺铂以及联合用药诱导细胞凋亡作用及细胞周期的影响将MCF-7用去甲斑蝥酸钠(30μg/ml、15μg/ml)处理48h后,G0/G1期细胞比例明显缩小,处于S期细胞比例增多,流式细胞检测结果表示人乳腺癌MCF-7细胞被去甲斑蝥酸钠阻滞在S期(p0.05),影响细胞无法向G2/M期转化,阻滞乳腺癌MCF-7细胞的脱氧核糖核酸合成,抑制肿瘤细胞的增殖进行。顺铂(1.25μg/ml、0.625μg/ml)作用于MCF-7细胞48h之后与对照组相比较,由细胞分布情况可见,G0/G1期细胞比例显著缩小,S期细胞比例明显增加,结果证明顺铂能够将人乳腺癌细胞增殖抑制在S期(p0.05),从而使细胞不能向G2/M期转化,进而实现对肿瘤细胞的增殖抑制作用。去甲斑蝥酸钠、顺铂联合作用于人乳腺癌MCF-7细胞的细胞凋亡率较单独用药组显著增加(p0.01)结论:1.去甲斑蝥酸钠、顺铂都可以抑制MCF-7细胞的增殖,在一定范围内,呈时间-浓度依赖性。2.同纯药组相比较,去甲斑蝥酸钠、顺铂两药联合后,对MCF-7细胞增殖抑制作用明显加强。3.去甲斑蝥酸钠、顺铂都可以使MCF-7细胞周期阻滞在S期,而联合用药组的细胞凋亡率显著高于单药组。
[Abstract]:Objective: to study the different concentrations of curcumin, sodium norcantharidate and cisplatin in different concentrations. The effects of monotherapy and combination on the proliferation inhibition, apoptosis induction and cell cycle of human breast cancer MCF-7 cell line were studied. The possible mechanism of sodium norcantharidate combined with cisplatin in inhibiting human breast cancer MCF-7 cell line was discussed. At the same time, the inhibition of cell proliferation was compared between single and combined drugs. So as to provide effective basic experimental data for breast cancer treatment strategy. Methods: 1. Human breast cancer MCF-7 cells were cultured to logarithmic growth phase in vitro. Curcumin at different concentrations was detected by MTT assay. Effects of sodium norcantharidate, cisplatin and two drugs on proliferation of MCF-7 cells after 24 hours and 72 hours. Flow cytometry was used to detect sodium norcantharidate 30 渭 g / ml 15 渭 g / ml, cisplatin 1.25 渭 g / ml / ml 0.625 渭 g / ml) alone and in combination with MCF-7 cells for 48 hours. Effects of apoptosis and cell cycle. Results: 1. Curcumin, curcumin, curcumin, curcumin, curcumin, curcumin, Inhibition of proliferation of MCF-7 cells by sodium norcantharidate and cisplatin. Sodium norcantharidate and cisplatin inhibited proliferation of MCF-7 cells in a time-concentration dependent manner. Curcumin had no obvious inhibitory effect on MCF-7 by statistical results. 2. The inhibitory effect of sodium norcantharidate and cisplatin on the proliferation of human breast adenocarcinoma MCF-7 cells combined with sodium norcantharidate 30 渭 g / ml 15 渭 g / ml and cisplatin 1.25 渭 g / ml 0.625 渭 g / ml. The inhibitory effects on human breast cancer MCF-7 cells were higher than those in the control group. With the increase of the concentration of norcantharidate and cisplatin, the inhibitory effect of sodium norcantharidate and cisplatin was enhanced. At the same time, the inhibitory effect of sodium norcantharidate and sodium norcantharidate on MCF-7 was also increased, and the inhibitory effect of sodium norcantharidate was enhanced with the prolongation of time. Apoptosis induced by cisplatin and combination of Cisplatin and its effect on Cell cycle the proportion of cells in G _ 0 / G _ 1 phase of MCF-7 treated with sodium norcantharidate 30 渭 g 路ml ~ (-1) 15 渭 g / ml for 48 h significantly decreased, and the proportion of cells in S phase increased. The results of flow cytometry showed that human breast cancer MCF-7 cells were blocked by sodium norcantharidate in S phase, which affected the cell transformation to G 2 / M phase, and blocked the synthesis of DNA in breast cancer MCF-7 cells. After treated with cisplatin 1.25 渭 g / ml, 0.625 渭 g / ml of cisplatin for 48 hours, compared with the control group, the proportion of cells in G _ 0 / G _ 1 phase decreased significantly and the proportion of cells in S phase increased significantly compared with the control group. The results showed that cisplatin could inhibit the proliferation of human breast cancer cells in S phase, thus the cells could not be transformed into G2 / M phase, and then the proliferation inhibition of tumor cells could be realized. The apoptotic rate of cisplatin combined with cisplatin on human breast cancer MCF-7 cells was significantly higher than that in the control group (P 0.01). Conclusion: 1. Sodium norcantharidate and cisplatin can inhibit the proliferation of MCF-7 cells. Compared with the pure drug group, the inhibitory effect of sodium norcantharidate and cisplatin on the proliferation of MCF-7 cells was significantly enhanced. 3. Sodium norcantharidate and cisplatin could block the cell cycle of MCF-7 in S phase. The apoptosis rate of the combined drug group was significantly higher than that of the single drug group.
【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】：R737.9

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