复方藤梨汤含药血清对人脑胶质瘤U251细胞增殖、凋亡及上皮间质转化的影响  

Effects of Fufang Tengli Decoction drug-containing serum on proliferation,apoptosis and epithelial-mesenchymal transition of human glioma U251 cells

作　　者：武海博 梁燕 沈雷 范展[1] 傅国惠 WU Haibo;LIANG Yan;SHEN Lei;FAN Zhan;FU Guohui(Department of Neurology,Nanyang Central Hospital,Nanyang 473000,China)

机构地区：[1]南阳市中心医院神经内科,473000

出　　处：《天津医药》2023年第8期841-846,共6页Tianjin Medical Journal

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20191452);河南省科技厅科技攻关项目(202102310408)。

摘　　要：目的探究复方藤梨汤含药血清对人脑胶质瘤U251细胞增殖、凋亡及上皮间质转化的影响。方法将24只雄性SD大鼠分为对照组和低、中、高剂量组[复方藤梨汤6、12、24g/(kg·d)],每组6只,制备含药血清处理人脑胶质瘤U251细胞。采用CCK-8法检测细胞活力,集落形成实验检测细胞克隆形成能力,流式细胞术检测细胞周期分布与细胞凋亡,定量聚合酶链反应检测原癌基因C-myc、细胞周期蛋白D1(CyclinD1)基因表达,Western blot法检测细胞凋亡蛋白[B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、胱天蛋白酶(Caspase)-3、Caspase-8]以及β-连环蛋白(β-catenin)、上皮性黏附蛋白(E-cadherin)及波形蛋白(Vimentin)表达。结果选取体积分数10%的含药血清为最终剂量。与对照组比较,低、中、高剂量组细胞活力、克隆形成数、C-myc mRNA、CyclinD1 mRNA、Vimentin表达水平降低,处于S期细胞比例、Bax/Bcl-2比值、Caspase-3表达水平升高(P<0.05);低、高剂量组G1期细胞比例降低(P<0.05);中、高剂量组G2期细胞比例、β-catenin表达水平降低,Caspase-8和E-cadherin表达水平升高(P<0.05);高剂量组细胞凋亡率升高(P<0.05)。结论复方藤梨汤含药血清可促使人脑胶质瘤U251细胞阻滞于S期,抑制细胞增殖,诱导细胞凋亡,通过调节β-catenin、E-cadherin和Vimentin表达抑制上皮间质转化。Objective To explore effects of Fufang Tengli Decoction drug-containing serum on the proliferation,apoptosis and epithelial-mesenchymal transition(EMT)of human glioma U251 cells.Methods A total of 24 male SD rats were divided into the control group,the low-dose,medium-dose and high-dose Fufang Tengli Decoction groups[6,12,24 g/(kg·d)],6 rats in each group.After 10 d of continuous intragastric administration,blood samples was collected to prepare drug-containing serum,and human glioma U251 cells were treated for 24 h.The cells activity was detected by Cell Counting Kit-8.The cell cloning was detected by colony formation assay.The distribution of cells cycles and apoptosis were detected by flow cytometry.The expressions of proto-oncogene C-myc and cyclin D1(CyclinD1)genes were detected by quantitative polymerase chain reaction.The expressions of B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),cysteine aspartase-3(caspase-3),caspase-8,β-catenin,epithelial cadherin(E-cadherin)and vimentin(Vimentin)were detected by Western blot assay.Results The drug-containing serum with 10%volume fraction was selected as the final dose.Compared with the control group,cells activity,number of clone formation and expression levels of C-myc,CyclinD1 mRNA and Vimentin were decreased,while numbers of cells in S phase and expression levels of Bax/Bcl-2 and Caspase-3 were increased in the low-dose,medium-dose and high-dose Fufang Tengli Decoction groups(P<0.05).The proportion of cells in G2 phase and expression level ofβ-catenin were decreased,while expression levels of Caspase-8 and E-cadherin were increased in the medium-dose and high-dose Fufang Tengli Decoction groups(P<0.05).Cell apoptosis rate was increased in the high-dose Fufang Tengli Decoction group(P<0.05).Conclusion Fufang Tengli Decoction drug-containing serum can block human glioma U251 cells in S phase,inhibit cell proliferation,induce apoptosis,regulate expressions ofβ-catenin,E-cadherin and Vimentin,and inhibit EMT.

关 键 词：神经胶质瘤 上皮-间质转化 细胞增殖 细胞凋亡 细胞增殖抑制药(中药) 复方藤梨汤含药血清 人脑胶质瘤U251细胞 

分 类 号：R739.41[医药卫生—肿瘤]