基于ROS治疗肿瘤的关键问题-ROS与其剂量效应的探讨
发布时间：2018-09-04 07:44
【摘要】：基于ROS治疗肿瘤是抗癌的一种新思路,目前被认为有可能成为肿瘤治疗的一种基本手段。与正常细胞相比,肿瘤细胞处于相对较高的氧化还原态,因此通过升高细胞ROS水平(促进ROS生成或/和抑制ROS清除)可使肿瘤细胞先于正常细胞达到ROS死亡阈值,进而死亡。在基于ROS的治疗假说中,ROS被认为是一种选择性杀伤肿瘤细胞的有效手段。剂量-效应关系是药物治疗学中最基本的治疗原则,而ROS剂量与细胞毒性之间是否存在剂量-效应关系,目前尚无报道。因此,我们开展该项研究进行ROS与其剂量效应关系的探讨。 在本研究中,我们随机选择了4种肿瘤细胞系(4T1、Bcap37、Hela、HepG2)和4例原代结直肠肿瘤细胞,利用不同氧化还原剂改变肿瘤细胞ROS水平,研究ROS改变对肿瘤细胞生长增殖的影响,进而分析ROS水平与其细胞毒性之间的相关性。结果发现,异硫氰酸苯乙酯(PEITC)和荜茇酰胺(PL)在升高肿瘤细胞ROS水平的同时导致细胞死亡；而与PEITC和PL相比,乳酸(LA)引起更高的ROS水平却不杀伤细胞；L-丁硫氨酸-亚砜亚胺(L-BSO)比PEITC、PL能更有效地降低细胞内GSH含量、更显著地升高ROS水平,却允许细胞进行性增殖。这些结果提示肿瘤细胞增殖或死亡与ROS水平高低或GSH含量多少之间无明显相关性。为进一步验证细胞死亡与ROS是否相关,我们通过还原剂氮乙酰半胱氨酸(NAC)和SOD-CAT类似物(EUK8、EUK134)降低PEITC、PL等药物升高的ROS后,观察细胞增殖或死亡情况。实验结果表明NAC (4mM)可逆转PEITC、多柔比星(Dox)或三氧化二砷(ATO)升高的ROS,但不能阻断它们导致的细胞死亡；而相反,NAC (4mM)不能逆转PL升高的ROS,却可完全阻断其导致的细胞死亡；EUK8、EUK134均可逆转上述药物升高的ROS,但均不能阻断药物导致的细胞死亡。这说明细胞死亡与否与ROS高低之间不存在相关性。最后,我们在原代结直肠肿瘤细胞中发现,LA在显著升高肿瘤细胞ROS水平的同时并不杀伤肿瘤细胞；而与LA相比,PEITC、PL或ATO升高的ROS水平相对较低,但均导致细胞死亡。 综上所述,本研究发现ROS水平与其细胞毒性之间无明显的剂量-效应关系,如果ROS是导致细胞死亡的直接原因,它就应该符合药物治疗中最基本的剂量-效应关系原则。这是基于ROS治疗肿瘤中亟待解决的关键问题。
[Abstract]:Tumor therapy based on ROS is a new way of anticancer, and it is considered to be a basic method of tumor therapy. Compared with normal cells, tumor cells are in a relatively high redox state. Therefore, by increasing the level of ROS (promoting ROS production or / and inhibiting ROS clearance), tumor cells can reach ROS death threshold before normal cells and then die. Ros is considered to be an effective method to selectively kill tumor cells in the treatment hypothesis based on ROS. The dose-effect relationship is the most basic therapeutic principle in drug therapeutics, but whether there is a dose-effect relationship between the dose of ROS and cytotoxicity has not been reported. Therefore, we carried out this study to investigate the relationship between ROS and its dose effect. In this study, we randomly selected four tumor cell lines (4T1 Bcap37 Hela HepG2) and four primary colorectal cancer cells to change the ROS level of tumor cells with different oxidizing reductants, and to study the effect of ROS changes on the growth and proliferation of tumor cells. The correlation between ROS level and cytotoxicity was analyzed. The results showed that ethyl isothiocyanate (PEITC) and Piper longum (PL) increased the ROS level of tumor cells and resulted in cell death, but lactic (LA) induced higher ROS level than PEITC and PL, but did not kill the cells. Compared with PEITC,PL, L-butylthiaminine-sulfoxide (L-BSO) can decrease the GSH content more effectively and increase the ROS level more significantly, but it allows the cells to proliferate progressively. These results suggest that there is no significant correlation between proliferation or death of tumor cells and the level of ROS or the content of GSH. To further verify the correlation between cell death and ROS, we observed cell proliferation or cell death after reducing the ROS of PEITC,PL by reducing PEITC,PL (NAC) and SOD-CAT analogue (EUK8,EUK134). The results showed that NAC (4mM) could reverse the cell death induced by PEITC, doxorubicin (Dox) or arsenic trioxide (ATO), but could not block the cell death caused by (ATO), whereas 4mM (4mM) could not reverse the PL elevation of ROS, but could completely block the cell death. EUK8,EUK134 could reverse the increase of ROS, but could not block the cell death caused by the drug. This suggests that there is no correlation between cell death and ROS. Finally, we found that the ROS level of primary colorectal tumor cells was significantly increased by LLA, while the level of ROS was lower than that of LA, but both of them resulted in cell death. In conclusion, there is no significant dose-effect relationship between ROS level and cytotoxicity. If ROS is the direct cause of cell death, it should conform to the most basic dose-effect relationship principle in drug therapy. This is a key problem to be solved in the treatment of tumor based on ROS.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R73-36
本文编号：2221438
[Abstract]:Tumor therapy based on ROS is a new way of anticancer, and it is considered to be a basic method of tumor therapy. Compared with normal cells, tumor cells are in a relatively high redox state. Therefore, by increasing the level of ROS (promoting ROS production or / and inhibiting ROS clearance), tumor cells can reach ROS death threshold before normal cells and then die. Ros is considered to be an effective method to selectively kill tumor cells in the treatment hypothesis based on ROS. The dose-effect relationship is the most basic therapeutic principle in drug therapeutics, but whether there is a dose-effect relationship between the dose of ROS and cytotoxicity has not been reported. Therefore, we carried out this study to investigate the relationship between ROS and its dose effect. In this study, we randomly selected four tumor cell lines (4T1 Bcap37 Hela HepG2) and four primary colorectal cancer cells to change the ROS level of tumor cells with different oxidizing reductants, and to study the effect of ROS changes on the growth and proliferation of tumor cells. The correlation between ROS level and cytotoxicity was analyzed. The results showed that ethyl isothiocyanate (PEITC) and Piper longum (PL) increased the ROS level of tumor cells and resulted in cell death, but lactic (LA) induced higher ROS level than PEITC and PL, but did not kill the cells. Compared with PEITC,PL, L-butylthiaminine-sulfoxide (L-BSO) can decrease the GSH content more effectively and increase the ROS level more significantly, but it allows the cells to proliferate progressively. These results suggest that there is no significant correlation between proliferation or death of tumor cells and the level of ROS or the content of GSH. To further verify the correlation between cell death and ROS, we observed cell proliferation or cell death after reducing the ROS of PEITC,PL by reducing PEITC,PL (NAC) and SOD-CAT analogue (EUK8,EUK134). The results showed that NAC (4mM) could reverse the cell death induced by PEITC, doxorubicin (Dox) or arsenic trioxide (ATO), but could not block the cell death caused by (ATO), whereas 4mM (4mM) could not reverse the PL elevation of ROS, but could completely block the cell death. EUK8,EUK134 could reverse the increase of ROS, but could not block the cell death caused by the drug. This suggests that there is no correlation between cell death and ROS. Finally, we found that the ROS level of primary colorectal tumor cells was significantly increased by LLA, while the level of ROS was lower than that of LA, but both of them resulted in cell death. In conclusion, there is no significant dose-effect relationship between ROS level and cytotoxicity. If ROS is the direct cause of cell death, it should conform to the most basic dose-effect relationship principle in drug therapy. This is a key problem to be solved in the treatment of tumor based on ROS.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R73-36
【参考文献】
相关期刊论文 前2条
1 ;Reactive oxygen species: A double-edged sword in oncogenesis[J];World Journal of Gastroenterology;2009年14期
2 ;Dicoumarol Alters Cellular Redox State and Inhibits Nuclear Factor Kappa B to Enhance Arsenic Trioxide-Induced Apoptosis[J];Acta Biochimica et Biophysica Sinica;2004年03期
，本文编号：2221438
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