β拉帕醌联合PI3K/mTOR抑制剂NVP-BEZ235抑制BGC-823细胞增殖和迁移  

β-Lapachone combined with NVP-BEZ235 inhibit proliferation and migration of BGC-823 gastric cancer cells

作　　者：崔雪莲 陈丽艳[1] 孙洁 王艺璇 林贞花[1,2] CUI Xuelian;CHEN Liyan;SUN Jie;WANG Yixuan;LIN Zhenhua(Department of Cancer Research Center;Department of Pathology, Medical College, Yanbian University, Yanji 133002, China)

机构地区：[1]延边大学肿瘤研究中心 [2]延边大学医学院病理学教研室,吉林延吉133002

出　　处：《细胞与分子免疫学杂志》2018年第2期129-135,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(61371067);长白山学者特聘教授奖励计划(2016001)

摘　　要：目的研究β拉帕醌(β-lapachone)与磷脂酰肌醇3激酶/哺乳动物雷帕霉素靶蛋白(PI3K/m TOR)抑制剂NVP-BEZ235联合用药对人胃癌BGC-823细胞增殖与迁移的影响及机制。方法将BGC-823细胞分为空白对照组、1μmol/Lβ-lapachone处理组、50 nmol/L NVP-BEZ235处理组及1μmol/Lβ-lapachone联合50 nmol/L NVP-BEZ235处理组。采用MTT法和平板克隆形成实验检测BGC-823细胞的增殖能力;Western blot法检测细胞增殖相关蛋白磷酸化蛋白激酶B(p-AKT)、磷酸化核因子κB(p-NF-κB)、磷酸化的细胞外信号调节激酶(p-ERK)及细胞周期蛋白D1(cyclin D1)的蛋白水平;划痕实验及TranswellTM迁移实验检测细胞迁移能力;Western blot法检测上皮-间质转化(EMT)相关标志物上皮钙黏素(E-cadherin)、波形蛋白(vimentin)、锌指转录因子Snail及β联蛋白(β-catenin)的蛋白水平。结果与β-lapachone处理组或NVP-BEZ235处理组相比,β-lapachone联合NVP-BEZ235处理组能更显著地抑制细胞的增殖和克隆形成能力,p-AKT、p-NF-κB、p-ERK及cyclin D1的蛋白表达下调最明显;在联合处理组对细胞迁移的抑制更明显,EMT相关标志物E-cadherin表达上调,vimentin、Snail及β-catenin表达下调最明显。结论β-lapachone联合NVP-BEZ235可有效抑制BGC-823胃癌细胞增殖,可能与p-AKT、p-NF-κB、p-ERK和cyclin D1表达降低有关;并通过调控EMT进程,抑制BGC-823细胞的迁移。Objective To investigate the effect and molecular mechanism of β-Iapachone combined with NVP-BEZ235 on the proliferation and migration of BGC-823 human gastric cancer cells. Methods BGC-823 cells were randomly divided into four groups： control group, 1 umol/L β-Iapachone group, 50 nmol/L NVP-BEZ235 group and 1 μmol/L β-Iapachone combined with 50 nmol/L NVP-BEZ235 group. The proliferation of cells was determined using the MTT assay and colony formation assay. The expression levels of proliferation-related proteins phosphorylated AKT （p-AKT）, phosphorylated NF-κB (p-NF-κB）, phosphorylated extracellular signal-regulated kinase （p-ERK） and cyclin D1 were detected by Western blotting. The migration of cells was measured by wound healing assay and TranswellTM migration assay. The expression levels of epithelial-mesenchymal transition （EMT） markers E-cadherin, vimentin, Snail and β-catenin were detected by Western blotting. Results Compared with 13-1apachone or NVP-BEZ235 treatment, the combination of β-Iapachone and NVP-BEZ235 showed more prominent inhibitory effect on the proliferation and colony formation of BGC-823 cells. The most effective suppression on the expressions of p-AKT, p-NF-κB, p-ERK and cyclin D1 was observed in the combination therapy.Combined treatment also showed more evident inhibitory effect on the migration of BGC-823 cells as compared with β-lapachone or NVP-BEZ235 treatment. The expression of E-cadherin was significantly up-regulated, but the expressions of vimentin, Snail and β-catenin were significantly down-regulated by the combined treatment. Conclusion β-1apachone combined with NVP-BEZ235 can effectively inhibit the proliferation of BGC-823 cells, which may be related to the down-regulation of p-AKT, p-NF-κB, p-ERK and cyclin D1. Moreover, the combined treatment can effectively suppress the migration of BGC-823 cells via modulating the EMT process.

关 键 词：β-lapachone NVP—BEZ235 细胞增殖 迁移 上皮-间质转化 BGC-823细胞 

分 类 号：R285.5[医药卫生—中药学] Q279[医药卫生—中医学]