番泻苷B抑制STAT3和ERK1/2活化对鼻咽癌CNE-2细胞生长、侵袭及裸鼠成瘤的影响  

Effects of sennoside B on growth,invasion and tumorigenesis of nasopharyngeal carcinoma CNE-2 cells via inhibiting STAT3 and ERK1/2 activation

作　　者：魏璐璐[1] 吉文伟 黄维平[1] WEI Lulu;JI Wenwei;HUANG Weiping(Ward 1 of Ear,Nose and Throat,Nanyang Central Hospital,Nanyang Henan 473000,China;Department of General Biliary Surgery,Nanyang Central Hospital,Nanyang Henan 473000,China)

机构地区：[1]河南省南阳市中心医院耳鼻喉一病区,河南南阳473000 [2]河南省南阳市中心医院胆道普外科,河南南阳473000

出　　处：《临床与病理杂志》2020年第3期547-555,共9页Journal of Clinical and Pathological Research

基　　金：河南省科技厅科技攻关项目(112102310173)。

摘　　要：目的:以鼻咽癌CNE-2细胞及其移植瘤小鼠模型为研究对象,探讨番泻苷B(Sennoside B)对鼻咽癌CNE-2细胞生长、侵袭及裸鼠成瘤的影响及其作用机制。方法:以5,10,20μmol/L处理CNE-2细胞后,采用BrdU染色法检测细胞增殖,Hoechst染色法检测细胞凋亡,Transwel l检测细胞侵袭,划痕试验检测细胞迁移,蛋白质印迹法检测Ki-67、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、半胱氨酸天冬氨酸蛋白酶-3(caspase-3)、caspase-9、血管内皮生长因子(vascular endothelial growth factor,VEGF)、E-钙黏蛋白(E-cadherin)和N-钙黏蛋白(N-cadherin)的表达情况及信号转导与转录激活因子3(signal transducer and activator of transcription 3,STAT3)和细胞外调节蛋白激酶1/2(extracellular regulated protein kinases 1/2,ERK1/2)的磷酸化情况;最后,建立荷瘤小鼠模型,分别腹腔注射25,50,100 mg/kg番泻苷B,30 d后检测肿瘤重量,并用免疫组织化学检测肿瘤组织中Ki-67和VEGF的表达。结果:体外实验示番泻苷B能剂量依赖性地降低BrdU阳性细胞率(P<0.05),抑制增殖蛋白Ki-67,PCNA表达(P<0.05),增加细胞凋亡率(P<0.05),上调凋亡蛋白caspase-3,caspase-9表达(P<0.05),抑制细胞侵袭和迁移能力(P<0.05),调节细胞上皮-间充质转化(epithelial mesenchymal transition,EMT)相关蛋白VEGF,E-cadherin和N-cadherin表达(P<0.05),并降低信号通路蛋白STAT3,ERK1/2的磷酸化水平(P<0.05)。体内实验示番泻苷B能明显减轻肿瘤重量(P<0.05),并下调肿瘤组织中Ki-67,VEGF的表达水平(P<0.05)。结论:番泻苷B通过抑制STAT3,ERK1/2的磷酸化激活,对鼻咽癌CNE-2细胞的生长、侵袭及裸鼠成瘤产生抑制作用。Objective:To investigate the effects of sennoside B on the growth,invasion and tumorigenesis of nasopharyngeal carcinoma CNE-2 cells.Methods:CNE-2 cells were treated with 5,10,20μmol/L;BrdU staining was used to detect cell proliferation;Hoechst staining was used to detect cell apoptosis;Transwell staining was used to detect cell invasion;Scratch test was used to detect cell migration;Western blot was used to detect the protein expression of Ki-67,PCNA,caspase-3,caspase-9,VEGF,E-cadherin and N-cadherin and the phosphorylation of STAT3 and ERK1/2.Finally,after establishing the tumor-bearing mice model,rats were respectively intraperitoneal injected 25,50 and 100 mg/kg sennoside B.The weight of tumors was measured at the day 30,and the expressions of Ki-67 and VEGF in tumors were detected by immunohistochemistry.Results:In vitro experiment:sennoside B could reduce BrdU positive cell rate in a dose-dependent manner(P<0.05),inhibit the expression of Ki-67 and PCNA(P<0.05),increase apoptotic rate(P<0.05),up-regulate the expression of caspase-3 and caspase-9(P<0.05),inhibit cell invasion and migration(P<0.05),regulate the expression of EMT related protein VEGF,E-cadherin and N-cadherin(P<0.05),and reduce the phosphorylation of STAT3,ERK1/2(P<0.05).In vivo experiments:sennoside B could significantly reduce the weight of tumors(P<0.05),and down-regulate the expression of Ki-67 and VEGF in tumors(P<0.05).Conclusion:Sennoside B inhibits the growth,invasion and tumorigenesis of nasopharyngeal carcinoma CNE-2 cells by inhibiting the activation of STAT3 and ERK1/2.

关 键 词：鼻咽癌 番泻苷B 信号转导与转录激活因子3 细胞外调节蛋白激酶 

分 类 号：R285.5[医药卫生—中药学]