成分分析

紫菀水提物对人肺癌A549细胞增殖、侵袭及裸鼠成瘤能力的影响及作用机制探讨*

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  • 1.长沙民政职业技术学院,长沙 410004;
    2.南华大学附属南华医院胸外科,衡阳 421002;
    3.南华大学护理学院基础护理教研室,衡阳 421001;
    4.南华大学附属南华医院重症医学科,衡阳 421002
第一作者 Tel:(0731)82804949; E-mail:yaopingbo752300@126.com
**Tel:(0731)82804949; E-mail:zjx92020@163.com

收稿日期: 2021-02-10

  网络出版日期: 2024-06-26

基金资助

*湖南省自然科学基金项目(2018JJ6122)

Effects and aster water extract on proliferation and invasion of human lung cancer A549 cells, and tumorigenesis ability of nude mice*

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  • 1. Changsha Civil Affairs Vocational and Technical College, Changsha 410004, China;
    2. Department of Thoracic Surgery, Nanhua Hospital Affiliated to Nanhua University, Hengyang 421002, China;
    3. Department of Basic Nursing, School of Nursing, Nanhua University, Hengyang 421001, China;
    4. Department of Critical Care Medicine, Nanhua Hospital Affiliated to Nanhua University, Hengyang 421002, China

Received date: 2021-02-10

  Online published: 2024-06-26

摘要

目的: 探讨紫菀水提物对人肺癌A549细胞增殖、侵袭及裸鼠成瘤能力的影响及作用机制。方法: 将体外培养MRC-5和A549细胞分别与不同浓度AWE(0、0.6、1.25、2.5、5、10、20、40、80、160、320 g·L-1)孵育,根据MTT实验设置A549细胞为AWE 0 g·L-1组、AWE 10 ·L-1组、AWE 20 ·L-1组和AWE 40 ·L-1组。CCK-8法检测各组细胞增殖,Transwell法检测侵袭细胞数。采用Wnt/β-catenin信号通路抑制剂XAV939处理A549细胞,Western blot检测各组细胞Wnt-1、β-catenin、血管内皮生长因子(VEGF)和Ki-67蛋白表达。皮下种植A549细胞悬液(5×106个·mL-1)0.2 mL,经AWE(灌胃5 g·kg-1·d-1,4周)干预后,进行移植瘤组织称重,免疫组化检测VEGF和Ki-67表达。结果: AWE对MRC-5细胞无明显抑制作用(P>0.05),对A549细胞的抑制作用呈药物浓度依赖性,IC50=24.81 g·L-1,随着AWE浓度增加,AWE浓度≥20 g·L-1时,细胞存活率下降(P<0.05)。AWE 20 g·L-1组和AWE 40 g·L-1组A549细胞增殖活性和侵袭细胞数低于AWE 0 g·L-1组(P<0.05)。AWE 40 g·L-1组和XAV939组A549细胞Wnt-1、β-catenin、Ki67和VEGF蛋白表达低于对照组(P<0.05);AWE 40 +XAV939组A549细胞Wnt-1、β-catenin、Ki67和VEGF蛋白表达低于AWE 40 g·L-1组和XAV939组(P<0.05)。裸鼠成瘤实验结果显示,AWE组裸鼠移植瘤质量低于对照组(P<0.05),且移植瘤组织中Ki67和VEGF蛋白表达低于对照组(P<0.05)。结论: AWE可以抑制肺癌细胞增殖和侵袭,抑制移植瘤生长,可能与抑制Wnt/β-catenin信号通路有关。

本文引用格式

姚平波, 刘雨露, 朱子贵, 赵红, 张建新 . 紫菀水提物对人肺癌A549细胞增殖、侵袭及裸鼠成瘤能力的影响及作用机制探讨*[J]. 药物分析杂志, 2022 , 42(3) : 380 -386 . DOI: 10.16155/j.0254-1793.2022.03.03

Abstract

Objective: To explore the effects and mechanisms of aster water extract (AWE) on the proliferation and invasion of human lung cancer A549 cells, and the tumorigenesis capability of nude mice. Methods: MRC-5 and A549 cells were cultured in vitro and incubated with different concentrations of AWE (0, 0.6, 1.25, 2.5, 5, 10, 20, 40, 80, 160, 320 g·L-1). In the MTT assay, A549 cells were divided into AWE 0 g·L-1 group, AWE 10 g·L-1 group, AWE 20 g·L-1 group and AWE 40 g·L-1 group, and the cell proliferation rate in each group was detected by CCK-8. The number of invasion cells was detected using Transwell method. A549 cells were treated with Wnt/β-catenin signaling pathway inhibitor XAV939. The expression of β-catenin, vascular endothelial growth factor (VEGF) and Ki-67 proteins in each group was detected by Western blot. About 0.2 mL of A549 cells suspension (5×106 cell·mL-1) was subcutaneously implanted. After the intervention of AWE (5 g·kg-1·d-1 intragastric administration, 4 weeks), transplanted tumor tissues was weighed. The expression of VEGF and Ki-67 was detected by immunohistochemistry. Results: There was no significant inhibitory effect of AWE on MRC-5 cells (P>0.05), the inhibitory effect on A549 cells showed concentration-dependence (IC50=24.81 g·L-1). With the increase of AWE concentration, when AWE concentration was not lower than 20 g·L-1, survival rate of cells was decreased (P<0.05). The proliferation activity of A549 cells and number of invasion cells in AWE 20 g·L-1 group and AWE 40 g·L-1 group were lower than those in AWE 0 g·L-1 group (P<0.05). The expressions of Wnt-1, β-catenin, Ki67 and VEGF proteins in A549 cells of AWE 40 g·L-1 group and XAV939 group were lower than those in control group (P<0.05), which were lower in AWE 40 g·L-1+XAV939 group than AWE 40 g·L-1 group and XAV939 group (P<0.05). The results of tumorigenicity assay in nude mouse showed that the mass of transplanted tumors in AWE group was lower than that in control group (P<0.05), and the expression of Ki67 and VEGF proteins in transplanted tumor tissue was lower than that in control group (P<0.05). Conclusion: AWE can inhibit the proliferation and invasion of lung cancer cells, and inhibit growth of transplanted tumors, which may be related to the inhibition the expression of Wnt/β-catenin signaling pathway.

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