全反式维甲酸对胶质瘤细胞增殖作用的影响

发布时间：2018-06-02 21:38

本文选题：全反式维甲酸 + 胶质瘤　；参考：《苏州大学》2014年硕士论文

【摘要】：胶质瘤是颅内肿瘤中发病率最高的，其发病机制尚未完全明确，主要的治疗方法是手术加放化疗，但并未明显提高患者的生存时间和生活质量。全反式维甲酸(Alltrans retinoic acid, ATRA)在急性早幼粒细胞白血病（acute promyelocytic leukemia，APL）的治疗中取得成功，其中ATRA是通过结合维甲酸受体（Retinoic acid receptor，RAR）发挥其作用，且ATRA对实体肿瘤的作用也逐渐被发现。但有关ATRA作用于胶质瘤的研究很少，因此有进一步研究的价值。目的：研究ATRA及维甲酸受体激动剂或拮抗剂对U251胶质瘤细胞增殖和迁移的影响，探讨维甲酸受体激动剂或拮抗剂对ATRA抑制U251细胞增殖的调节，为ATRA能作为胶质瘤治疗的有效药物提供研究依据。方法：(1)使用1、2.5、5、10、20μMATRA分别处理U251细胞48h，观察ATRA对U251细胞的形态学变化和细胞迁移作用的影响，MTT检测ATRA对细胞的增殖作用的影响；(2)10μM ATRA分别联合四种药物[维甲酸受体激动剂(RARα激动剂，BMS7531μM)；维甲酸受体拮抗剂(RARα拮抗剂， Ro41-52531μM)；维甲酸受体激动剂(RXRα激动剂，CD32541μM)：④维甲酸受体（RXRα拮抗剂，UVI30031μM）]处理U251细胞48h，观察对U251细胞增殖和迁移的影响，MTT检测各组对细胞的增殖作用的影响；(3) Western blot检测U251细胞RARa，RXRa的表达情况。结果：(1)细胞划痕和MTT显示，ATRA可以抑制U251细胞的增殖与迁移，并呈剂量依赖性，ATRA联合RARα激动剂BMS753与ATRA组相比，对U251细胞增殖迁移的抑制作用增强；ATRA联合RARα拮抗剂Ro41-5253与ATRA组相比，对U251细胞增殖迁移的抑制作用减弱。(2) Western blot检测结果显示ATRA可以提高RARα受体蛋白表达，而对RXRα受体蛋白的作用不明显，ATRA联合RARα激动剂BMS753与ATRA组相比， RARα受体蛋白表达上升；ATRA联合RARα拮抗剂Ro41-5253组与ATRA组相比，RARα受体蛋白表达下降；ATRA联合RXRα激动剂CD3254与ATRA组相比，RXRα受体蛋白表达上升；ATRA联合RXRα拮抗剂UVI3003与ATRA组相比，RXRα受体蛋白表达量下降。结论：上述结果表明，ATRA对U251细胞增殖及迁移有抑制作用，这种抑制作用与RARα受体密切相关；RARα激动剂BMS753可以增强ATRA对U251细胞的抑制作用，RARα拮抗剂Ro41-5253可以减弱ATRA对U251细胞的抑制作用，这种作用与RARα受体表达水平相关联。研究结果为ATRA用于胶质瘤治疗的可行性及临床应用中联合用药以减少ATRA的使用量、降低其副作用的发生提供了依据。
[Abstract]:Glioma is the highest incidence of intracranial tumors, its pathogenesis is not completely clear, the main treatment is surgery plus radiotherapy and chemotherapy, but has not significantly improved the survival time and quality of life of patients. All trans retinoic acid, ATRA) is successful in the treatment of acute promyelocytic leukemia (APL), in which ATRA plays its role by binding retinoic acid receptor, Retinoic acid receptor rarr, and the effect of ATRA on solid tumors is gradually discovered. However, there are few studies on the effect of ATRA on glioma, so it is valuable to further study. Aim: to investigate the effects of ATRA and retinoic acid receptor agonists or antagonists on the proliferation and migration of U251 glioma cells, and to investigate the effects of ATRA agonists or antagonists on the proliferation of U251 glioma cells. To provide the basis for the study of ATRA as an effective drug for the treatment of glioma. Methods 1) U251 cells were treated with 10 渭 MATRA for 48 hours. The morphological changes of ATRA on U251 cells and the effect of cell migration were observed. The effects of ATRA on proliferation of U251 cells were detected by MTT. The effects of 10 渭 M ATRA combined with four kinds of drugs [retinoic acid receptor] on the proliferation of U251 cells were determined. U251 cells were treated with RXR 伪 agonist (BMS 7531 渭 M), retinoic acid receptor antagonist (Ro41-52531 渭 MN), RXR 伪 agonist (CD32541 渭 M) and RXR 伪 antagonist (UVI 30031 渭 M) for 48 h. The effects of RXR 伪 antagonist on proliferation and migration of U251 cells were observed for 48 h. The expression of RARARXRa in U251 cells was detected by Western blot. Results the cell scratches and MTT showed that ATRA could inhibit the proliferation and migration of U251 cells in a dose-dependent manner, and the inhibitory effect of ATRA combined with RAR 伪 agonist BMS753 on the proliferation and migration of U251 cells was enhanced compared with that of ATRA group and RAR 伪 antagonist Ro41-5253 group. The inhibitory effect on proliferation and migration of U251 cells was attenuated. The results of Western blot analysis showed that ATRA could increase the expression of RAR 伪 receptor protein. But the effect on RXR 伪 receptor protein was not obvious. The expression of RAR 伪 receptor protein increased in ATRA group and RAR 伪 antagonist Ro41-5253 group compared with ATRA group. The expression of RAR 伪 receptor protein in ATRA + RXR 伪 agonist CD3254 and ATRA group was lower than that in ATRA group. Compared with ATRA group, the expression of RXR 伪 receptor protein was increased. The expression of RXR 伪 receptor protein in ATRA combined with RXR 伪 antagonist UVI3003 was lower than that in ATRA group. Conclusion: these results suggest that ATRA can inhibit the proliferation and migration of U251 cells. This inhibitory effect is closely related to RAR 伪 receptor. BMS753 can enhance the inhibitory effect of ATRA on U251 cells. RAR 伪 antagonist Ro41-5253 can attenuate the inhibitory effect of ATRA on U251 cells, which is related to the expression level of RAR 伪 receptor. The results provide a basis for the feasibility of ATRA in the treatment of glioma and the clinical application of combined use of ATRA to reduce the amount of ATRA and to reduce the occurrence of side effects.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.41

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