雷公藤甲素对胶质瘤的影响

Impacts of triptolide on glioma

目的通过体内实验和体外实验两个水平来研究雷公藤甲素对胶质瘤治疗的影响,其中体外实验采取一般给药及纳米载药两种方式进行研究,雷公藤甲素对胶质瘤的影响是否可能是通过抑制mir-221的表达而产生。方法在纳米给药的基础上,体内实验和体外实验两个水平来研究,应用细胞计数试剂盒(CCK-8)、Transwell、细胞划痕实验方法检测各实验组中细胞增殖迁移能力的变化;采用蛋白质免疫印迹(Western blot)检测各实验组中核因子κB(NF-κB),磷酸化核因子(pNF-κB)以及第10号染色体缺失的磷酸酶及张力蛋白同源的基因(PTEN)三种蛋白的表达。结果CCK-8实验发现随着药物作用时间延长和药物浓度增加,细胞存活率逐渐下降;Transwell细胞迁移实验,发现雷公藤甲素对C6细胞的迁移有明显的抑制作用,特别是中浓度组和高浓度组;细胞划痕实验结果和Transwell细胞迁移实验得出的结论一致;Western blot检测结果为各组中NF-κB蛋白的表达量由高到低分别是:纳米用药组、普通用药组、生理盐水组,纳米用药组的PTEN蛋白表达高于普通用药组和生理盐水组;实时定量PCR技术,检测发现雷公藤甲素对mir-221的表达量有抑制作用。结论雷公藤甲素可能是通过抑制NF-κB的磷酸化从而减少mir-221的激活进而上调抑癌基因PTEN蛋白的表达而产生影响,而且雷公藤甲素纳米载药的效果可能更好。此外雷公藤甲素可以抑制C6细胞的生长和迁移以及mir-221的表达。

Objective To investigate the impacts of triptolide in the treatment of glioma at the levels of in vivo and in vitro experiments.The in vitro experiments were carried out in two ways,which were general administration and nano-drug loading,to investigate whether the impacts of triptolide on glioma were caused by inhibiting the expression of mir-221.Methods On the basis of nano-drug administration,two levels,namely in vivo experiment and in vitro experiment,were investigated.The changes of cell proliferation and migration ability in each of the experimental group were detected by using cell counting kit(CCK-8),Transwell and cell scratch test.Protein Western blot was used to detect the expression of three kinds of proteins of the nuclear factorκB(NF-κB),the phosphorylated nuclear factor(pNF-κB)and the phosphatase and tensin homolog deleted on chromosome ten(PTEN)in each of the experimental group.Results CCK-8 experiment showed that the cell survival rate decreased gradually with the prolongation of drug action time and the increase of drug concentration;Transwell cell migration experiment showed that triptolide significantly inhibited the migration of C6 cells,especially in the medium and high concentration groups;The result of cell scratch test was consistent with that of Transwell cell migration experiment;The results of Western blot detection showed that the ranking of the expression of NF-κB protein in each group from high to low was the nano-drug administration group,the general administration group and the normal saline group.The expression of PTEN protein in the nano-drug administration group was higher than that in the general adm inistration group and the nor mal saline group;Real-time quantitative PCR technology showed that triptolide inhibited the expression of mir-221.Conclusion Triptolide may have an impact by inhibiting the phosphorylation of NF-κB,thus reducing the activation of mir-221 and then up-regulating the expression of cancer suppressor gene PTEN protein.Moreover,the effect of triptolide nano-drug loading may be better.In addition,triptolide can inhibit the growth and migr ation of C6 cells and the expression of mir-221.