本文選題：香豆素 + 輻射增敏　； 參考：《北京協(xié)和醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:香豆素類(lèi)衍生物具有良好的抗腫瘤活性,但線粒體靶向香豆素類(lèi)化合物未見(jiàn)報(bào)道。本文工作合成了攜帶不同靶向基團(tuán)的線粒體靶向香豆素,并對(duì)其抗腫瘤作用和輻射增敏作用進(jìn)行了研究。方法:通過(guò)將6-甲基香豆素溴化,隨后將溴化后的產(chǎn)物與靶向基團(tuán)連接得到線粒體靶向的香豆素。利用MTT法檢測(cè)非靶向香豆素以及不同靶向基團(tuán)的線粒體靶向香豆素對(duì)腫瘤細(xì)胞的抑制作用以及對(duì)正常細(xì)胞A31細(xì)胞的毒性�？寺⌒纬蓪�(shí)驗(yàn)考察非靶向香豆素以及不同靶向基團(tuán)的線粒體靶向香豆素對(duì)細(xì)胞增殖的影響,采用多靶單擊擬合模型方程計(jì)算(4-(二甲氨基)吡啶)甲基香豆素放射增敏比,評(píng)價(jià)增敏效果。DCFH-DA法檢測(cè)非靶向香豆素以及不同靶向基團(tuán)的線粒體靶向香豆素對(duì)腫瘤細(xì)胞ROS水平的影響。Rh123法檢測(cè)非靶向香豆素以及不同靶向基團(tuán)的線粒體靶向香豆素對(duì)腫瘤細(xì)胞線粒體膜電勢(shì)的影響。Rh-2-AM法檢測(cè)非靶向香豆素以及以三苯基膦為靶向基團(tuán)的線粒體靶向香豆素對(duì)腫瘤細(xì)胞線粒體Ca2+濃度的影響。Hoechst 33258染色法測(cè)定非靶向香豆素以及不同靶向基團(tuán)的線粒體靶向香豆素對(duì)腫瘤細(xì)胞凋亡的影響。NAO染色法測(cè)定非靶向香豆素以及以三苯基膦為靶向基團(tuán)的線粒體靶向香豆素對(duì)腫瘤細(xì)胞線粒體質(zhì)量的影響。結(jié)果:通過(guò)質(zhì)譜,核磁共振氫譜和碳譜,確認(rèn)了目標(biāo)化合物結(jié)構(gòu)。MTT結(jié)果顯示,不同靶向基團(tuán)的線粒體靶向香豆素能有效降低腫瘤細(xì)胞的存活率,在50-500μmol/L濃度范圍內(nèi)對(duì)正常細(xì)胞A31無(wú)明顯毒性,而非靶向香豆素對(duì)腫瘤細(xì)胞存活率無(wú)明顯影響�？寺⌒纬蓪�(shí)驗(yàn)表明不同靶向基團(tuán)的線粒體靶向香豆素都可以有效抑制腫瘤細(xì)胞的增殖,而非靶向香豆素對(duì)腫瘤細(xì)胞增殖無(wú)明顯影響。結(jié)果表明6-(4-(二甲氨基)吡啶)甲基香豆素化合物濃度為IC20時(shí)的輻射增敏比1.58,ROS實(shí)驗(yàn)結(jié)果表明,和照射組相比,6-(4-(二甲氨基)吡啶)甲基香豆素明顯提高了 A549細(xì)胞中的ROS水平。而6-(煙酰胺)甲基香豆素和6-(三苯基膦)甲基香豆素,和對(duì)照組相比,則明顯提高了腫瘤細(xì)胞內(nèi)的ROS水平。而非靶向香豆素對(duì)腫瘤細(xì)胞內(nèi)的ROS水平無(wú)明顯影響。Rh123實(shí)驗(yàn)結(jié)果表明,與對(duì)照組相比,6-(煙酰胺)甲基香豆素和6-(三苯基膦)甲基香豆素有效地降低了腫瘤細(xì)胞的線粒體膜電勢(shì)。而非靶向香豆素對(duì)腫瘤細(xì)胞線粒體膜電勢(shì)無(wú)明顯影響。線粒體Ca2+測(cè)定實(shí)驗(yàn)結(jié)果表明,6-(三苯基膦)甲基香豆素可以明顯提高HeLa細(xì)胞中線粒體Ca2+濃度。而非靶向香豆素對(duì)HeLa細(xì)胞中線粒體Ca2+濃度無(wú)明顯影響。Hoechst 33258染色結(jié)果表明,6-(煙酰胺)甲基香豆素和6-(三苯基膦)甲基香豆素可以誘導(dǎo)腫瘤細(xì)胞產(chǎn)生凋亡。非靶向香豆素不能有效誘導(dǎo)HeLa細(xì)胞凋亡。NAO染色實(shí)驗(yàn)結(jié)果表明,6-(三苯基膦)甲基香豆素有效地降低了HeLa細(xì)胞的線粒體質(zhì)量。非靶向香豆素對(duì)HeLa細(xì)胞中線粒體質(zhì)量無(wú)明顯影響。結(jié)論:非靶向香豆素對(duì)HeLa細(xì)胞存活率,增殖,凋亡無(wú)明顯抑制效果。6-(4-(二甲氨基)吡啶)甲基香豆素可以明顯地提高A549細(xì)胞對(duì)放射線的敏感性。6-(煙酰胺)甲基香豆素通過(guò)提高ROS水平和降低線粒體膜電勢(shì),促使腫瘤細(xì)胞凋亡。6-(三苯基膦)甲基香豆素則可以通過(guò)提高腫瘤細(xì)胞中ROS水平、降低線粒體膜電勢(shì)、提高線粒體Ca2+濃度和降低線粒體質(zhì)量,促使腫瘤細(xì)胞凋亡。因此通過(guò)將非靶向香豆素設(shè)計(jì)成線粒體靶向藥物可顯著提高其抗腫瘤活性。
[Abstract]:Objective: coumarin derivatives have good antitumor activity, but mitochondria targeted coumarins have not been reported. In this paper, the mitochondrial targeting coumarin with different target groups was synthesized, and its anti-tumor effect and radiation sensitization were studied. Method: bromination of 6- methyl coumarin and then bromine The transformed product was linked with the target group to obtain the mitochondrial targeting coumarin. The inhibitory effect of the mitochondrial targeting coumarin on the tumor cells and the toxicity to the normal cell A31 cells were detected by the MTT method. The cloning and formation of the non targeting coumarin and the lines of different target groups were investigated. The effect of marinin on the proliferation of cells was measured by a multi target clicking model equation (4- (two methylamino) pyridine) methyl coumarin radiosensitization ratio, and the effect of.DCFH-DA method to detect the effect of non targeting coumarin and the mitochondrial target of different target groups on the ROS level of tumor cells by.Rh123 method was not detected. Effects of coumarin and different targeting groups targeting coumarin on the mitochondrial membrane potential of tumor cells.Rh-2-AM detection of non targeting coumarin and mitochondrial target of three phenyl phosphine as the target group on the concentration of mitochondrial Ca2+ in tumor cells by the.Hoechst 33258 staining method for the determination of non targeting coumarin The effect of coumarin on the apoptosis of tumor cells with different target groups,.NAO staining method was used to determine the effect of coumarin on the mitochondrial quality of tumor cells by targeting coumarin and the target group with three phenyl phosphine as the target group. Results: the target compound structure.MT was confirmed by mass spectrometry, nuclear magnetic resonance hydrogen spectrum and carbon spectrum. T results showed that the mitochondrial targeting coumarin of different target groups could effectively reduce the survival rate of tumor cells, and there was no obvious toxicity to normal cell A31 in the concentration range of 50-500 mu mol/L, but the non targeting coumarin had no significant effect on the survival rate of tumor cells. The proliferation of tumor cells was inhibited effectively, but the non targeting coumarin had no obvious effect on the proliferation of tumor cells. The results showed that the radiation sensitization ratio of 6- (4- (two methylamino) pyridine) methyl coumarin compound was 1.58. The results of ROS experiment showed that 6- (4- (two methylamino) pyridine) methyl coumarin increased A5 obviously compared with the irradiated group. The level of ROS in 49 cells. While 6- (nicotinamide) methylene coumarin and 6- (three phenyl phosphine) methyl coumarin, compared with the control group, significantly increased the level of ROS in the tumor cells. The non targeting coumarin did not significantly affect the ROS level in the tumor cells, and the.Rh123 experiment showed that the 6- (nicotinamide) methyl coumarin and 6- (nicotinamide) were compared with the control group. Three phenyl phosphine) methyl coumarin effectively reduced the mitochondrial membrane potential of tumor cells. Non targeting coumarin had no significant effect on the mitochondrial membrane potential of tumor cells. Mitochondrial Ca2+ assay showed that 6- (three phenyl phosphine) methylene coumarin could significantly increase the mitochondrial Ca2+ concentration in HeLa cells, but not targeted coumarin to HeLa The results of.Hoechst 33258 staining showed that 6- (nicotinamide) methylene coumarin and 6- (three phenyl phosphine) methyl coumarin could induce apoptosis in tumor cells. Non targeting coumarin could not effectively induce HeLa cell apoptosis.NAO staining experimental results showed that 6- (three phenyl phosphine) methyl coumarin was reduced effectively. The mitochondrial mass of HeLa cells was lower. The non targeting coumarin had no significant effect on the mitochondrial mass in HeLa cells. Conclusion: non targeting coumarin has no obvious inhibitory effect on the survival rate, proliferation and apoptosis of HeLa cells.6- (4- (two methylamino) pyridine) methyl coumarin can significantly increase the sensitivity of A549 cells to radiation sensitivity.6- (nicotinamide) methyl. Coumarin can increase ROS level and decrease mitochondrial membrane potential, and induce apoptosis of tumor cells.6- (three phenyl phosphine) methylene coumarin, which can improve the mitochondrial Ca2+ concentration and decrease the mitochondrial mass and induce apoptosis by improving the ROS level in tumor cells, so that the non targeting coumarin is designed. Mitochondrial targeting drugs can significantly improve its antitumor activity.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R914;R96

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 王華偉;王華南;康肖夢(mèng);張宇睿;楊福軍;徐文清;;線粒體靶向抗氧化劑MitoQ的抗腫瘤效應(yīng)[J];輻射研究與輻射工藝學(xué)報(bào);2016年01期

2 Massimo Donadelli;Ilaria Dando;Elisa Dalla Pozza;Marta Palmieri;;Mitochondrial uncoupling protein 2 and pancreatic cancer:A new potential target therapy[J];World Journal of Gastroenterology;2015年11期

3 劉樹(shù)森;;線粒體呼吸鏈與活性氧[J];生命科學(xué);2008年04期

4 江穎娟;曾耀英;王通;肇靜嫻;刑飛躍;王希朝;梁佩燕;;喜樹(shù)堿誘導(dǎo)Jurkat細(xì)胞線粒體膜電勢(shì)和線粒體質(zhì)量變化的研究[J];中國(guó)病理生理雜志;2006年05期

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