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
| 作者: |
1魏璐璐,
2吉文伟,
1黄维平
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 |
| 通讯: |
魏璐璐
Email: nvtig0@sina.com |
| DOI: | 10.3978/j.issn.2095-6959.2020.03.003 |
| 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5，10，20 μmol/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，Transwell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。
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
Effects of sennoside B on growth, invasion and tumorigenesis of nasopharyngeal carcinoma CNE-2 cells via inhibiting STAT3 and ERK1/2 activation
CorrespondingAuthor: WEI Lulu Email: nvtig0@sina.com
DOI: 10.3978/j.issn.2095-6959.2020.03.003
Foundation: This work was supported by the Key Technological Research Projects of Henan Provincial Science and Technology Department, China (112102310173).
Abstract
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, 20 μmol/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.
Keywords：
nasopharyngeal carcinoma; sennoside B; signal transducer and activator of transcription 3; extracellular regulated protein kinase 1/2