ERK介导姜黄素抑制人脑胶质瘤细胞U251侵袭性

Curcumin Inhibits the Tumor Invasiveness Via ERK/MAPK Pathway in Human Glioma U251 Cells

关键词:  姜黄素  胶质瘤  ERK  侵袭  免疫印迹

Key words:  curcumin  glioma  ERK  invasiveness  Western blot

摘要:

目的 探讨姜黄素对人脑胶质瘤细胞U251侵袭性的影响及ERK/MAPK信号通路在此过程中的作用。方法 将人脑胶质瘤细胞U251分为对照组和药物组，对照组以含10％小牛血清的DMEM培养基常规培养，药物组用16 μmol·L-1姜黄素（本实验先前的研究结果显示，姜黄素作用于U251细胞48 h的半数抑制剂量为16 μmol·L-1）处理。 Transwell侵袭小室模型法检测细胞侵袭性的变化；免疫细胞化学法检测基质金属蛋白酶－9（MMP-9）蛋白表达变化；免疫印迹（Western blot）法检测细胞外调节蛋白激酶（ERK）和磷酸化细胞外调节蛋白激酶（pERK）表达变化。结果 Transwell侵袭小室模型法显示，在每个高倍镜视野下，对照组穿过微孔膜的细胞数平均值为160个，药物组为42个。免疫细胞化学法显示姜黄素作用后MMP-9蛋白阳性细胞百分率由76％降低至40％。Western blot法显示，16 μmol·L-1姜黄素分别作用于U251细胞0、24、48和96 h，ERK蛋白水平无明显改变，pERK水平与ERK水平比值(pERK/ERK) 分别为0.48、0.42、0.35和0.22。结论 姜黄素在体外可有效抑制胶质瘤U251细胞的侵袭性，其机制可能是通过抑制异常激活的ERK信号通路来实现的。

Abstract:

OBJECTIVETo investigate the inhibitory effect of curcumin on the invasiveness of human glioma U251 cells in vitro and the role of ERK/MAPK pathway in this process. METHODSWe studied on the two groups:the control group and the treatment group with curcumin in U251 cells. Transwell invasion assay was used to measure the invasive ability of U251 cells. Expression of MMP-9 was detected by immunocytochemistry. The levels of total and phosphorylated ERK were assessed by Western blot analysis. RESULTSTranswell invasion assay showed that the number of cells through the basal membrane decreased from 160 to 42 per high vision.The immunocytochemistry experiment showed that the percentage of MMP-9 positive cells decreased from 76% to 40% after treatment.In the Western blot analysis, when the curcumin concentration came to 16 μmol·L-1 and the time of therapy differed,which was 0，24，48 or 96 h,the ratio of pERK to ERK was 0.48，0.42，0.35 and 0.22. CONCLUSIONCurcumin can inhibit the invasiveness of human glioma U251 cells in vitro. The inhibitory effect may result from the blocking of ERK pathway.