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羽扇豆醇通过MAPKs信号通路对人乳腺癌MCF-7细胞增殖的抑制作用研究
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| 篇名： | 羽扇豆醇通过MAPKs信号通路对人乳腺癌MCF-7细胞增殖的抑制作用研究 |
| TITLE： | |
| 摘要： | 目的：探讨羽扇豆醇对人乳腺癌MCF-7细胞增殖的抑制作用及可能机制。方法：以MCF-7细胞为对象，采用MTT法检测不同剂量羽扇豆醇（7.5、15、30、60、90 mg/L）作用24 h后的细胞增殖情况，并计算细胞存活率和半数抑制浓度（IC50）；采用倒置显微镜和细胞克隆试验分别观察和检测不同剂量羽扇豆醇（15、30、60 mg/L）作用24 h后的细胞形态学特征以及克隆集落形成情况，并计算克隆形成率；分别采用MTT法和Western blotting法检测加用丝裂原活化蛋白激酶（MAPKs）信号通路相关调控蛋白抑制剂（PD98059、SP600125、SB203580）后细胞的增殖情况和相关调控蛋白[细胞外调节蛋白激酶1/2（ERK1/2）、磷酸化ERK1/2（p-ERK1/2）、c-Jun氨基末端激酶（JNK）、磷酸化JNK（p-JNK）、p38丝裂原活化蛋白激酶（p38 MAPK）、磷酸化p38 MAPK（p-p38 MAPK）]的表达情况。结果：经15、30、60、90 mg/L羽扇豆醇作用后，细胞的存活率均显著降低（P＜0.05或P＜0.01）；该化合物的IC50值为52.94 mg/L。经15、30、60 mg/L羽扇豆醇作用后，各组细胞形态均有所改变，并可见细胞脱落、漂浮、固缩、变圆、体积变小、坏死等现象，其克隆集落形成均有所减少，克隆形成率均显著降低（P＜0.05或P＜0.01）。单用羽扇豆醇时，细胞的存活率（60 mg/L羽扇豆醇）均较对照组显著降低，p-ERK1/2（15、30、60 mg/L羽扇豆醇）、p-JNK（30、60 mg/L羽扇豆醇）、p-p38 MAPK（30、60 mg/L羽扇豆醇）的相对表达量均较对照组显著升高（P＜0.05或P＜0.01）；加用相应抑制剂后，联用组细胞的存活率均较60 mg/L羽扇豆醇组显著升高，p-ERK1/2、p-JNK、p-p38 MAPK的相对表达量均较60 mg/L羽扇豆醇组显著降低（P＜0.05或P＜0.01）。结论：羽扇豆醇对人乳腺癌MCF-7细胞的增殖具有明显的抑制作用，其机制可能与促进MAPKs信号通路相关调控蛋白的磷酸化有关。 |
| ABSTRACT： | OBJECTIVE： To investigate inhibitory effects of lupeol on the proliferation of human breast cancer MCF-7 cells and its possible mechanism. METHODS： Taking MCF-7 cells as research object， MTT assay was used to detect the proliferation of MCF-7 cells after treated with different doses of lupeol （7.5， 15， 30， 60， 90 mg/L） for 24 h. Survival rate and IC50 of MCF-7 cells were calculated. The inverted microscope and cell cloning experiment were used to observe and detect the morphological characteristics of MCF-7 cells and clonal colony formation after treated with different doses of lupeol （15， 30， 60 mg/L） for 24 h. The rate of clonal colony formation was calculated. MTT method and Western blotting assay were used to detect the proliferation of MCF-7 cells and the expression of related regulatory proteins （ERK1/2， p-ERK1/2， JNK， p-JNK， p38 MAPK， p-p38 MAPK） after additionally treated with MAPKs signaling pathway-related regulation protein inhibitors PD98059， SP600125 and SB203580. RESULTS： After treated with 15， 30， 60， 90 mg/L lupeol， survival rates of MCF-7 cells were decreased significantly （P＜0.05 or P＜0.01）. IC50 value of the compound was 52.94 mg/L. After treated with 15， 30， 60 mg/L lupeol， the morphological characteristics of cells in each group changed， and the phenomena of cell exfoliation， floating， solid shrinkage， roundness， volume reduction and necrosis were observed. The formation of clonal colony decreased and the rate of clonal colony formation decreased significantly （P＜0.05 or P＜0.01）. When lupeol used alone， compared with control group， survival rate （60 mg/L lupeol）of MCF-7 cells was decreased significantly； the expression of p-ERK1/2 （15， 30， 60 mg/L lupeol）， p-JNK （30， 60 mg/L lupeol） and p-p38 MAPK （30， 60 mg/L lupeol） were increased significantly （P＜0.05 or P＜0.01）. After additional use of relevant inhibitors， compared with 60 mg/L lupeol group， survival rates of MCF-7 cells in combination groups were increased significantly， while relative expression of p-ERK1/2， p-JNK and p-p38 MAPK were decreased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： Lupeol can inhibit the proliferation of human breast cancer MCF-7 cells， the mechanism of which may be related to the phosphorylation of MAPKs signaling pathway-related regulatory proteins. |
| 期刊： | 2019年第30卷第14期 |
| 作者： | 江兴菊，潘年松，田晓云，黄梅，罗俊 |
| AUTHORS： | JIANG Xingju，PAN Niansong，TIAN Xiaoyun，HUANG Mei，LUO Jun |
| 关键字： | 羽扇豆醇；乳腺癌；MCF-7细胞；增殖；丝裂原活化蛋白激酶信号通路；调控蛋白；磷酸化 |
| KEYWORDS： | Lupeol； Breast cancer； MCF-7 cells； Proliferation； MAPKs signaling pathway； Regulatory protein； Phosphorylation |
| 阅读数： | 257 次 |
| 本月下载数： | 3 次 |
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