淫羊藿苷激活抑制鼻咽癌CNE-2细胞存活、侵袭、迁移的体内外研究  

Icariin Inhibits Survival,Invasion and Migration of Nasopharyngeal Carcinoma CNE-2 Cells in Vitro and in Vivo

作　　者：魏璐璐[1] 吉文伟 黄维平[1] WEI Lu-Lu;JI Wen-Wei;HUANG Wei-Ping(Inpatient Ward NO.1 of Otolaryngology,Nanyang Central Hospital,Nanyang 473000 Henan,China;Dept,of General Biliary Surgery,Nanyang Central Hospital,Nanyang 473000 Henan,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：《广州中医药大学学报》2021年第6期1223-1229,共7页Journal of Guangzhou University of Traditional Chinese Medicine

基　　金：河南省科技厅科技攻关项目(编号:112102310173)。

摘　　要：【目的】探讨淫羊藿苷对鼻咽癌CNE-2细胞存活和侵袭迁移特性的影响。【方法】(1)体外研究:用不同浓度淫羊藿苷处理鼻咽癌CNE-2细胞,采用细胞计数试剂盒8(CCK8)测定细胞增殖能力以选择合适的剂量进行后续实验。采用Transwell实验观察细胞侵袭能力;划痕实验观察细胞迁移能力;蛋白免疫印迹法检测血管内皮生长因子(VEGF)、上皮钙黏附蛋白(E-cadherin)、神经性钙黏附蛋白(N-cadherin)、波形蛋白(Vimentin)表达情况及信号通路蛋白Ca2+/钙调蛋白依赖性的蛋白激酶Ⅱ(CaMKⅡ)和c-Jun氨基末端激酶(JNK)的磷酸化情况。并进一步观察单独或联合JNK抑制剂SP600125对JNK活化程度的影响,采用蛋白免疫印迹法检测细胞JNK磷酸化水平。(2)体内研究:建立鼻咽癌CNE-2荷瘤裸鼠模型,测定裸鼠移植瘤质量、细胞凋亡和移植瘤组织VEGF表达情况、MVD值。【结果】选择低细胞毒性10、20、50μmol/L剂量的淫羊藿苷进行后续实验。与空白对照组(淫羊藿苷0μmol/L)比较,淫羊藿苷10、20、50μmol/L组CNE-2细胞侵袭细胞数、划痕愈合率降低,细胞VEGF、N-cadherin、Vimentin表达水平明显降低(P <0.05),E-cadherin表达水平与CaMKⅡ、JNK的磷酸化水平升高(P <0.05),均呈剂量依赖性。淫羊藿苷(10μmol/L)可减弱JNK抑制剂SP600125对JNK磷酸化的抑制作用。与模型组比较,淫羊藿苷组鼻咽癌裸鼠存活率升高,移植瘤质量减小,移植瘤组织细胞凋亡率升高,VEGF表达水平、MVD值明显下降(P <0.05)。【结论】淫羊藿苷可促进CaMKⅡ/JNK通路激活来抑制鼻咽癌CNE-2细胞的存活、侵袭和迁移能力。Objective To explore the effects of icariin on survival, invasion and migration of nasopharyngeal carcinoma CNE-2 cells. Methods(1)In vitro:Nasopharyngeal carcinoma CNE-2 cells were treated with different concentrations of icariin, and then cell count kit 8(CCK8)assay was used to detect cells viability to select appropriate dosages for subsequent experiment. The cell invasion ability was observed by Transwell assay,cell migration ability was observed by scratch-wound assay, expression levels of vascular endothelial growth factor(VEGF), E-cadherin, N-cadherin, Vimentin and phosphorylation levels of Ca2+/calcium-and calmodulin-dependent protein kinase Ⅱ(CaMKⅡ)and c-Jun N-terminalkinase(JNK)were detected by Western blotting assay. Furthermore, to observe the effects of JNK inhibitor SP600125 alone or combination on JNK activation degrees,the phosphorylation level of JNK was detected by Western blotting assay.(2)In vivo:a nasopharyngeal carcinoma CNE-2-bearing nude mouse model was established. The xenograft mass,apoptosis and expression of VEGF, and microvascular density(MVD) in Xenografts from nude mice were measured. Results The concentrations(10, 20, 50 μmol/L) with low cytotoxicity of icariin were selected for subsequent experiment.Compared with the blank control group(icariin of 0 μmol/L),the number of migrating cells and scratch-wound healing rate were decreased in the icariin 10,20,50 μmol/L groups,expression levels of VEGF,N-cadherin,Vimentin in cells were decreased,and expression level of E-cadherin and phosphorylation levels of CaMKⅡ and JNK in cells were increased(all P < 0.05),all the effects being in dose-dependent mode. Icariin had weakened the inhibitory effect of JNK inhibitor SP600125 on JNK phosphorylation level. Compared with the model group,the xenograft mass,rate of cell apoptosis,expression level of VEGF and MVD in Xenografts were up-regulated in the icariin group(all P < 0.05). Conclusion Icariin is effective for inhibiting the survival,invasion and migration of nasopharyngeal carcinom

关 键 词：淫羊藿苷 鼻咽癌 Ca2+/钙调蛋白依赖性的蛋白激酶Ⅱ C-JUN氨基末端激酶 CNE-2细胞 裸鼠 

分 类 号：R285.5[医药卫生—中药学]