大黄酸通过抑制ERK途径诱导胶质瘤细胞增殖分化的实验研究

Experimental study of rhein on glioma cell proliferation and differentiation by inhibiting ERK pathway

目的探讨大黄酸对胶质瘤细胞增殖和分化的影响,以及其可能的机制。方法大鼠F98和人Hs683细胞系分别予以不同浓度的大黄酸处理(大黄酸处理组)和单用培养基培养(对照组)。采用四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)比色法检测各组细胞的增殖能力;倒置显微镜观察细胞形态变化;免疫细胞化学检测细胞胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)的表达。ELISA法检测经25μmol/L大黄酸处理不同时间的细胞磷酸化细胞外信号调节激酶(phospho-excelluar signal-regulated kinase,p-ERK)的活性。结果与对照组相比,大黄酸处理组F98细胞的相对活性降低,细胞胞体失去了对照组细胞的多边形形状,胞体变小,并发出较长的突起;免疫细胞化学检测发现GFAP表达增加。大黄酸处理后的F98细胞p-ERK相对活性降低。用ERK1/2特异性抑制剂PD98059处理的F98细胞及25μmol/L大黄酸处理的人Hs683细胞增值活性下降,细胞形态出现与大黄酸处理F98细胞相似的变化,GFAP表达亦明显增加。结论大黄酸抑制胶质瘤细胞增殖,并可能通过抑制ERK途径促进胶质瘤细胞向成熟星形胶质细胞分化。

Objective To explore the effect of rhein on glioma cell proliferation and differentiation by inhibiting ERK pathway.Methods Rats glioma cell line F98 and human glioma cell line Hs683 were used in the study.The experiment was divided into a blank control group and a rhein-treated experimental group.MTT assay was used to determine the cell proliferation ability.Inverted microscope was used to observe the cellular morphological changes.Immunocytochemistry was used to detect Glial fibers acidic protein（GFAP） expression.ELISA method was used to detect p-ERK activity.Results In the rhein experimental group,the relative proliferation activities were decreased.Glioma cells monitored under microscope brought a major change to their structure morphology.The shape of rhein treated F98 glioma cells were analogous to that of matured astrocytes with tiny cell bodies and considerably extended,fine tapering systems.Immunocytochemistry results exhibited an indicative increase of GFAP expression in rhein treated cells.The relative p-ERK activities of F98 were decreased after treated by rhein.The relative p-ERK activities decreased and GFAP expression increased and morphology changed for F98 cells treated by PD98059 the specific ERK1/2 inhibitor and Hs683 cells treated by 25 μmol/L rhein.These changes were similar to the F98cells change after being treated by rhein.Conclusion Rhein may inhibit glioma cell proliferation and promote differentiation by inhibiting ERK pathway.