Effect of Astragalus Polysaccharide on Radiosensitivity and Epithelial-mesenchymal Transition of Human Nasopharyngeal Carcinoma CNE-1 Cells

Shu-cong ZHANG; Zhi-xiang CAI; Xue-tao WANG; Zhi-ying LI; Hui-sheng SONG

doi:10.13422/j.cnki.syfjx.20201967

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Effect of Astragalus Polysaccharide on Radiosensitivity and Epithelial-mesenchymal Transition of Human Nasopharyngeal Carcinoma CNE-1 Cells

Pharmacology | 更新时间：2020-09-16

- Effect of Astragalus Polysaccharide on Radiosensitivity and Epithelial-mesenchymal Transition of Human Nasopharyngeal Carcinoma CNE-1 Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 20, Pages: 59-66(2020)
- 作者机构：
  
  1.广州中医药大学第二临床医学院，广州 50006
  2.广州市番禺中心医院，广州 511400
  3.广州中医药大学第二附属医院，广州 510006
  4.广州医科大学第六附属医院清远市人民医院，广东清远 511518
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20201967
  CLC： R22;R242;R2-031;R285.5
- Received：05 January 2020，
  
  Published Online：27 July 2020，
  
  Published：20 October 2020
- 稿件说明：
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张树聪,蔡治祥,王学涛等.黄芪多糖对人鼻咽癌CNE-1细胞的放疗增敏及上皮间质转化的作用[J].中国实验方剂学杂志,2020,26(20):59-66.

ZHANG Shu-cong,CAI Zhi-xiang,WANG Xue-tao,et al.Effect of Astragalus Polysaccharide on Radiosensitivity and Epithelial-mesenchymal Transition of Human Nasopharyngeal Carcinoma CNE-1 Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(20):59-66.
张树聪,蔡治祥,王学涛等.黄芪多糖对人鼻咽癌CNE-1细胞的放疗增敏及上皮间质转化的作用[J].中国实验方剂学杂志,2020,26(20):59-66. DOI： 10.13422/j.cnki.syfjx.20201967.

ZHANG Shu-cong,CAI Zhi-xiang,WANG Xue-tao,et al.Effect of Astragalus Polysaccharide on Radiosensitivity and Epithelial-mesenchymal Transition of Human Nasopharyngeal Carcinoma CNE-1 Cells[J].Chinese Journal of Experimental Traditional Medical Formulae,2020,26(20):59-66. DOI： 10.13422/j.cnki.syfjx.20201967.

摘要

目的

采用黄芪多糖（APS)与放射治疗（IR）联用，研究APS对人鼻咽癌CNE-1细胞的放疗敏感性及上皮间质转化（EMT）的作用机制。

方法

采用细胞计数（CCK-8）法检测不同质量浓度APS（0，6.25，12.5，25，50，100，200 g·L

-1

）对CNE-1细胞的细胞毒性；克隆形成实验计算12.5 g·L

-1

APS与不同放射剂量（0，2，4，6 Gy）联用后对CNE-1细胞的存活分数（SF），利用线性二次方程数学模型（LQ）根据SF值绘制放射敏感曲线；细胞划痕和transwell小室实验分别检测各组细胞的迁移和侵袭能力；流式细胞仪检测各组细胞的凋亡情况；蛋白免疫印迹法（Western blot）检测各组细胞的EMT标记物、凋亡标记物以及蛋白激酶B/细胞外调节蛋白激酶（Akt/ERK）通路蛋白的表达水平。

结果

克隆形成实验和放射敏感曲线结果表明，非毒性剂量12.5 g·L

-1

APS与4 Gy放射剂量联合给药可以明显增加CNE-1细胞的放疗敏感性；与空白组及IR组比较，APS与IR联用可以抑制CNE-1细胞的迁移和侵袭能力（

0.05），明显增加CNE-1细胞的凋亡率（

0.05）。与空白组及IR组比较，APS与IR联用可以显著下调间质型钙黏蛋白（N-cadherin），p-Akt和p-ERK蛋白表达水平，明显上调上皮型钙黏蛋白（E-cadherin），B淋巴细胞瘤-2相关X蛋白（Bax）和半胱天冬氨酸蛋白酶-3（Caspase-3）蛋白表达水平（

0.05）。

结论

APS与IR联用可以抑制CNE-1细胞的迁移和侵袭，增加放疗引起的细胞凋亡，其可能通过抑制EMT和Akt/ERK通路有关。

Abstract

Objective

Astragalus polysaccharide (APS) was used in combination with ionizing radiation (IR) to investigate the mechanism of APS on the radiosensitivity of human nasopharyngeal narcinoma CNE-1 cells and the epithelial-mesenchymal transition (EMT).

Method

Cell counting kit-8 (CCK-8) was used to detect the cytotoxicity of different concentrations of APS (0，6.25，12.5，25，50，100，200 g·L

-1

) on CNE-1 cells. Colony formation assay was used to calculate the survival fraction (survival fraction

SF) of CNE-1 cells treated with 12.5 g·L

-1

APS combined with different radiation doses (0，2，4，6 Gy). The linear quadratic equation mathematical model (LQ) was used to draw the radiosensitivity curve according to SF value. Cell scratch and transwell chamber test were used to detect the migration and invasion ability of cells in each group. The apoptosis of cells in each group was detected by flow cytometry

Western blot was used to detect the expressions of EMT markers

apoptosis markers and protein kinase B/extracellular regulated protein kinases (Akt/ERK) pathway proteins in each group.

Result

The results of colony formation assay and radiosensitivity curve showed that the combination of non-toxic dose of 12.5 g·L

-1

APS and radiation dose of 4 Gy could significantly increase the radiosensitivity of CNE-1 cells. Compared with blank group and IR group

APS combined with IR could significantly inhibit the migration and invasion of CNE-1 cells (

0.05)

and increase the rate of apoptosis (

0.05). In addition

compared with the blank group and the IR group

APS combined with IR could significantly down-regulate the expressions of N-cadherin

p-Akt and p-ERK

and significantly up-regulate the expressions of E-cadherin

Bax and Caspase-3 (

0.05).

Conclusion

APS combined with IR can inhibit the migration and invasion of CNE-1 cells

and increase the apoptosis induced by radiotherapy

which may be related to the inhibition of EMT and Akt/ERK pathway.

关键词

Keywords

references

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