尼莫地平注射液抑制ERK1/2和P38 MAPK的磷酸化对人星形胶质瘤细胞U-251 MG生长和运动的抑制作用  

Inhibitory Effect of Nimodipine Injection on Growth and Motor Ability of Human Astroglioma Cell U-251 MG by Inhibiting Phosphorylation of ERK1/2 and p38 MAPK

作　　者：潘冬 罗卢 粟珏佳 程远 PAN Dong;LUO Lu;SU Juejia;CHENG Yuan(Department of Pharmacy,Sichuan Armed Police Corps Hospital,Leshan,Sichuan,614000,China;Department of Health Service,Sichuan Armed Police Corps Hospital,Leshan,Sichuan,614000,China;Department of Administrative,Sichuan Armed Police Corps Hospital,Leshan,Sichuan,614000,China;Department of Information,Hospital of the 74 th Group Army,Guangzhou,510000,China)

机构地区：[1]武警四川省总队医院药剂科,四川省乐山市614000 [2]武警四川省总队医院卫勤处,四川省乐山市614000 [3]武警四川省总队医院行政处,四川省乐山市614000 [4]解放军第七十四集团军医院信息科,广州市510000

出　　处：《医学分子生物学杂志》2021年第4期291-297,共7页Journal of Medical Molecular Biology

摘　　要：目的采用体外培养人星形胶质瘤U-251MG细胞为研究对象,探讨尼莫地平注射液(Nimodip-ine,Nim)对其生长和运动的影响及其机制.方法CCK8法检测不同浓度Nim处理U-251MG后细胞存活率,选择无明显毒性的3个剂量进行后续实验.将细胞随机分为对照组、Nimodipine 0.125 mg组、Nimodip-ine 0.25 mg组和Nimodipine 0.5 mg组,采用Edu染色检测细胞增殖;Hoechst检测细胞凋亡情况;Transwell检测细胞侵袭;划痕实验检测细胞迁移;镜下观察细胞形态变化;Western印迹检测增殖相关蛋白Ki67、增殖细胞核抗原(PCNA),凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶3(caspase-3)、caspase-9,上皮间质转换(EMT)相关蛋白上皮细胞钙粘蛋白(E-cadherin)、神经钙粘蛋白(N-cadherin)、波形蛋白(Vimentin)的表达水平及信号通路细胞外信号调节激酶1/2(ERK1/2)、P38丝裂原活化蛋白酶(P38MAPK)的磷酸化水平.结果当Nim浓度达到1 mg/L时,可以对U-251MG细胞产生明显的细胞毒性,故选择无明显细胞毒作用的0.125、0.25和0.5 mg/L 3个剂量进行后续试验.Nim能剂量依赖性的抑制体外培养U-251MG细胞的增殖、侵袭和迁移能力,增加细胞凋亡率;促进E-cadherin,抑制caspase-3、caspase-9、Vimentin、N-cadherin、p-ERK1/2和p-P38的蛋白表达.结论Nim能有效调节人星形胶质瘤U-251MG细胞的生长和运动能力,其机制与调节ERK1/2和P38MAPK信号通路的磷酸化有关.Objective To investigate the effects of Nimodipine(Nim)on the growth and movement of human astroglioma U-251MG cells cultured in vitro and its mechanism.Methods CCK8 assay was used to detect the cell viability of U-251MG treated with different concentrations of Nim.Three doses without obvious toxicity were selected for subsequent experiments.The cells were randomly divided into control group,Nimodipine group(0.125 mg),Nimodipine group(0.25 mg)and Nimodipine group(0.5 mg).Cell proliferation was detected by Edu staining;apoptosis was detected by Hoechst;cell invasion was detected by Transwell;cell migration was detected by scratch test;cell morphology was observed by microscopy;proliferation-related proteins Ki67,proliferating cell nuclear antigen(PCNA),apoptosis-related proteins Cysteine aspartic protease 3(Caspase-3),caspase-9,Epithelial-mesenchymal transition(EMT)-related proteins Epithelial cell cadherin(E-cadherin),Neurocadherin(N-cadherin),Vimentin and phosphorylation levels of Extracellular signal-regulated kinase 1/2(ERK1/2)and P38 Mitogen-activated protein kinase(p38 MAPK)signaling pathways were detected by Western blotting.Results When the concentration of Nimodipineinjection reached 1 mg/L,it could induce obvious cytotoxicity to U-251 MG cells.Therefore,the doses of 0.125,0.25 and 0.5 mg/L were selected for following test.Nimodipine injection could inhibit the proliferation,invasion and migration of U-251MG cells in vitro in a dose-dependent manner,increase the apoptosis rate,promote E-cadherin,inhibit the expression of caspase-3,caspase-9,Vimentin,N-cadherin,extracellular signal-regulated ki-nasel/2(p-ERKl/2)and P38 Mitogen-activated protein kinase(P38MAPK)pro­teins.Conclusion Nimodipine injection can effectively regulate the growth and motility of human astroglioma U-251MG cells.Its mechanism is related to the regulation of phosphorylation of ERK1/2 and P38MAPK signaling pathways.

关 键 词：星形胶质瘤 尼莫地平 细胞外信号调节激酶1/2 P38丝裂原活化蛋白酶 

分 类 号：R739.41[医药卫生—肿瘤]