藁本内酯调节RhoA/ROCK信号通路对食管癌细胞生物学行为的影响  

Effects of ligustilide regulating RhoA/ROCK signaling pathway on biological behavior of esophageal cancer cells

作　　者：郝凯凯 王晓敏 刘峥[1] 刘东洋 李静[1] HAO Kaikai;WANG Xiaomin;LIU Zheng;LIU Dongyang;LI Jing(Department of Oncology,Handan Central Hospital,Hebei Province,Handan 056000,China;Department of Imaging,Affiliated Hospital of Hebei Engineering University;Department of Thoracic Surgery,Handan Central Hospital,Hebei Province)

机构地区：[1]河北省邯郸市中心医院肿瘤科,056000 [2]河北工程大学附属医院影像科 [3]河北省邯郸市中心医院胸外科

出　　处：《天津医药》2024年第11期1164-1170,共7页Tianjin Medical Journal

基　　金：河北省2022年度医学科学研究计划(20220505)。

摘　　要：目的探讨藁本内酯(LIG)对食管癌细胞增殖、凋亡、血管生成拟态及Ras同源基因家族蛋白A(Rho A)/Rho关联含卷曲螺旋结合蛋白激酶(ROCK)信号通路的影响。方法用浓度为0、12.5、25、50、100、200μmol/L LIG处理食管癌细胞EC-109,检测细胞活性,筛选适宜浓度进行后续实验。将EC-109细胞分为对照组(Control组),LIG低、中、高浓度组(LIG-L、LIG-M、LIG-H组),LIG高浓度+RhoA激活剂Naciclasine组(LIG-H+Naciclasine组)。Edu检测细胞增殖,流式细胞术检测细胞凋亡;观察血管生成拟态;Western blot检测细胞增殖、凋亡相关蛋白及RhoA、ROCK蛋白表达,裸鼠移植瘤实验验证LIG对食管癌肿瘤生长的影响,免疫组化检测移植瘤血管内皮生长因子(VEGF)、RhoA、ROCK表达水平。结果与Control组相比,LIG-L、LIG-M、LIG-H组EC-109细胞血管拟态管状结构依次减少,Edu阳性率、细胞周期蛋白(Cyclin)D1、细胞增殖核抗原(Ki67)、B细胞淋巴瘤/白血病-2(Bcl-2)、RhoA、ROCK表达依次降低,P21、细胞凋亡率、Bcl-2相关蛋白(Bax)、胱天蛋白酶(Caspase)-3表达依次升高(P<0.05)。RhoA激活剂Naciclasine可部分逆转LIG对食管癌细胞增殖、凋亡和血管生成拟态的改善作用。裸鼠移植瘤实验显示,与Control组相比,LIG组裸鼠移植瘤生长减缓,肿瘤体积减小,RhoA、ROCK、VEGF表达水平降低(P<0.05)。结论LIG通过抑制RhoA/ROCK信号通路抑制食管癌细胞的增殖及血管生成拟态,促进食管癌细胞凋亡。Objective To investigate effects of ligustilide(LIG)on proliferation,apoptosis,angiogenic mimicry and Ras homolog gene family member A(RhoA)/Rho associated coiled coil containing protein kinase 1(ROCK)signaling pathway in esophageal cancer cells.Methods Esophageal cancer cell line EC-109 was treated with LIG at concentrations of 0,12.5,25,50,100,and 200μmol/L to detect cell activity,and the suitable concentration was selected for subsequent experiments.EC-109 cells were grouped into the control group,the LIG low,medium and high concentration groups(LIG-L,LIG-M and LIG-H groups),and the LIG-H+RhoA activator Naciclassine group(LIG-H+Naciclassine group).Edu was applied to detect cell proliferation,and flow cytometry was applied to detect cell apoptosis.Angiogenetic mimicry was observed.Western blot assay was applied to detect expression levels of proteins related to cell proliferation and apoptosis,and RhoA,ROCK proteins.Nude mouse tumor transplantation experiment was applied to verify the effect of LIG on the growth of esophageal cancer tumors.Immunohistochemistry was applied to detect expression levels of angiogenesis related factors(VEGF),RhoA and ROCK proteins in transplanted tumors.Results Compared with the control group,the vascular mimicry tubular structure of EC-109 cells decreased sequentially in the LIG-L group,the LIG-M group and the LIG-H group.The positive rate of Edu,the expression levels of Cyclin D1,Ki67,Bcl-2,RhoA and ROCK reduced in turn.P21,cell apoptosis rate,the expression of Bax and Caspase-3 increased in sequence(P<0.05).Naciclasine,RhoA activator,partially reversed the effect of LIG on cell proliferation,apoptosis and vasculogenic mimicry of esophageal cancer cells.Nude mouse transplantation tumor experiment showed that compared with the control group,the growth rate of transplanted tumor showed down,tumor volume decreased and the expression levels of RhoA,ROCK and VEGF decreased in the LIG group(P<0.05).Conclusion Ligustilide inhibits the proliferation and angiogenic mimicry of esophageal c

关 键 词：藁本内酯 食管肿瘤 ρA GTP结合蛋白质 RHO相关激酶类 细胞增殖 细胞凋亡 肿瘤移植 

分 类 号：R735.1[医药卫生—肿瘤]