兩種抗癌中藥與順鉑聯(lián)用對人乳腺癌MCF-7細(xì)胞系增殖抑制作用及機(jī)制的研究
本文選題:去甲斑蝥酸鈉 切入點(diǎn):順鉑 出處:《吉林大學(xué)》2017年碩士論文 論文類型:學(xué)位論文
【摘要】:目的:本研究通過研究不同濃度的傳統(tǒng)中藥姜黃素、去甲斑蝥酸鈉和化療藥物順鉑在不同時(shí)間,單藥以及聯(lián)合用藥對人乳腺癌MCF-7細(xì)胞增殖抑制的作用以及誘導(dǎo)凋亡和細(xì)胞周期的影響,初步探討去甲斑蝥酸鈉與順鉑聯(lián)用抑制人乳腺癌MCF-7細(xì)胞株可能的作用機(jī)制,同時(shí)對比單獨(dú)用藥與聯(lián)合用藥細(xì)胞增殖抑制作用的情況,從而為乳腺癌治療策略提供有效的基礎(chǔ)實(shí)驗(yàn)數(shù)據(jù)做參考。方法:1、人乳腺癌細(xì)胞MCF-7細(xì)胞體外培養(yǎng)至對數(shù)生長期。2、通過MTT法檢測不同濃度的姜黃素、去甲斑蝥酸鈉和順鉑以及兩藥聯(lián)合作用于MCF-7細(xì)胞24、48和72小時(shí)后細(xì)胞增殖的情況。3、使用流式細(xì)胞儀檢測去甲斑蝥酸鈉(30μg/ml、15μg/ml)、順鉑(1.25μg/ml、0.625μg/ml)單獨(dú)以及聯(lián)合作用于MCF-7細(xì)胞48小時(shí)后誘導(dǎo)凋亡的情況和細(xì)胞周期的影響。結(jié)果:1.姜黃素、去甲斑蝥酸鈉和順鉑對MCF-7細(xì)胞增殖的抑制情況去甲斑蝥酸鈉和順鉑對MCF-7細(xì)胞均有增殖抑制作用,并且抑制效果呈時(shí)間-濃度依賴性的表現(xiàn),姜黃素通過實(shí)驗(yàn)的統(tǒng)計(jì)結(jié)果來看對MCF-7無明顯抑制作用。2.不同濃度去甲斑蝥酸鈉與順鉑聯(lián)用對人乳腺腺癌MCF-7細(xì)胞增殖的抑制作用聯(lián)合應(yīng)用去甲斑蝥酸鈉(30μg/ml、15μg/ml)和順鉑(1.25μg/ml、0.625μg/ml),對人乳腺癌MCF-7細(xì)胞的抑制作用均高于單獨(dú)用藥組,隨著去甲斑蝥酸鈉和順鉑的濃度增加,抑制作用增強(qiáng),同時(shí),單純的增加去甲斑蝥酸鈉的濃度,對MCF-7的抑制作用也在增強(qiáng),隨著作用時(shí)間的延長抑制作用增強(qiáng)。3.去甲斑蝥酸鈉、順鉑以及聯(lián)合用藥誘導(dǎo)細(xì)胞凋亡作用及細(xì)胞周期的影響將MCF-7用去甲斑蝥酸鈉(30μg/ml、15μg/ml)處理48h后,G0/G1期細(xì)胞比例明顯縮小,處于S期細(xì)胞比例增多,流式細(xì)胞檢測結(jié)果表示人乳腺癌MCF-7細(xì)胞被去甲斑蝥酸鈉阻滯在S期(p0.05),影響細(xì)胞無法向G2/M期轉(zhuǎn)化,阻滯乳腺癌MCF-7細(xì)胞的脫氧核糖核酸合成,抑制腫瘤細(xì)胞的增殖進(jìn)行。順鉑(1.25μg/ml、0.625μg/ml)作用于MCF-7細(xì)胞48h之后與對照組相比較,由細(xì)胞分布情況可見,G0/G1期細(xì)胞比例顯著縮小,S期細(xì)胞比例明顯增加,結(jié)果證明順鉑能夠?qū)⑷巳橄侔┘?xì)胞增殖抑制在S期(p0.05),從而使細(xì)胞不能向G2/M期轉(zhuǎn)化,進(jìn)而實(shí)現(xiàn)對腫瘤細(xì)胞的增殖抑制作用。去甲斑蝥酸鈉、順鉑聯(lián)合作用于人乳腺癌MCF-7細(xì)胞的細(xì)胞凋亡率較單獨(dú)用藥組顯著增加(p0.01)結(jié)論:1.去甲斑蝥酸鈉、順鉑都可以抑制MCF-7細(xì)胞的增殖,在一定范圍內(nèi),呈時(shí)間-濃度依賴性。2.同純藥組相比較,去甲斑蝥酸鈉、順鉑兩藥聯(lián)合后,對MCF-7細(xì)胞增殖抑制作用明顯加強(qiáng)。3.去甲斑蝥酸鈉、順鉑都可以使MCF-7細(xì)胞周期阻滯在S期,而聯(lián)合用藥組的細(xì)胞凋亡率顯著高于單藥組。
[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.
【學(xué)位授予單位】:吉林大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2017
【分類號】:R737.9
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