吉林大学学报(医学版)2024,Vol.50Issue(1) :25-32.DOI:10.13481/j.1671-587X.20240104

白屈菜红碱对人卵巢癌SKOV3细胞迁移、侵袭和上皮-间质转化的影响

Effect of chelerythrine on migration,invasion,and epithelial-mesenchymal transition of human ovarian cancer SKOV3 cells

周佳 邱智东 林喆 律广富 许佳明 林贺 王可欣 王雨辰 黄晓巍
吉林大学学报(医学版)2024,Vol.50Issue(1) :25-32.DOI:10.13481/j.1671-587X.20240104

白屈菜红碱对人卵巢癌SKOV3细胞迁移、侵袭和上皮-间质转化的影响

Effect of chelerythrine on migration,invasion,and epithelial-mesenchymal transition of human ovarian cancer SKOV3 cells

周佳 1邱智东 1林喆 1律广富 2许佳明 1林贺 1王可欣 1王雨辰 1黄晓巍3
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作者信息

  • 1. 长春中医药大学药学院临床药学与中药药理教研室,吉林 长春 130117
  • 2. 长春中医药大学吉林省人参科学研究院中药药理组,吉林 长春 130117
  • 3. 长春中医药大学药学院临床药学与中药药理教研室,吉林 长春 130117;长春中医药大学东北亚中医药研究院基础研究所,吉林 长春 130117
  • 折叠

摘要

目的:探讨白屈菜红碱(CHE)对人卵巢癌SKOV3细胞迁移、侵袭和上皮-间质转化(EMT)的抑制作用,阐明其相关作用机制.方法:体外培养SKOV3细胞,分为对照组和2.5、5.0、10.0、20.0及40.0 μmol·L-1 CHE组,采用噻唑蓝(MTT)法检测各组细胞增殖抑制率.体外培养SKOV3细胞,分为对照组、转移生长因子β1(TGF-β1)组、TGF-β1+5 μmol·L-1 CHE组和TGF-β1+10 μmol·L-1 CHE组,采用细胞划痕实验检测各组细胞迁移率,Transwell小室实验检测各组细胞中迁移细胞数和侵袭细胞数,Westren blotting法检测各组细胞中E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)蛋白表达水平,免疫荧光染色法检测各组细胞中E-cadherin和N-cadherin荧光强度.结果:MTT法,与对照组比较,5.0、10.0、20.0和 40.0 μmol·L-1 CHE组细胞增殖抑制率明显升高(P<0.05或P<0.01).细胞划痕实验,与对照组比较,TGF-β1组细胞迁移率明显升高(P<0.01);与TGF-β1组比较,TGF-β1+5 μmol·L-1 CHE组和 TGF-β 1+ 10 μmol·L-1 CHE组细胞迁移率明显降低(P<0.01).Transwell小室实验,与对照组比较,TGF-β1组细胞中迁移细胞数和侵袭细胞数明显增加(P<0.05);与TGF-β1 组比较,TGF-β1+5 μmol·L-1 CHE组和TGF-β1+10 μmol·L-1 CHE组细胞中迁移细胞数和侵袭细胞数明显减少(P<0.01).Westren blotting法,与对照组比较,TGF-β1组细胞中E-cadherin蛋白表达水平明显降低(P<0.01),N-cadherin和Vimentin蛋白表达水平明显升高(P<0.05或P<0.01);与TGF-β1组比较,TGF-β1+ 5 μmol·L-1 CHE组和TGF-β1+10 μmol·L-1 CHE组细胞中E-cadherin蛋白表达水平明显升高(P<0.01),N-cadherin和Vimentin蛋白表达水平明显降低(P<0.01).免疫荧光染色,与对照组比较,TGF-β1组细胞中E-cadherin荧光强度明显降低,N-cadherin荧光强度明显升高;与TGF-β1组比较,TGF-β1+5 μmol·L-1 CHE组和TGF-β1+10 μmol·L-1 CHE组细胞中E-cadherin荧光强度明显升高,N-cadherin荧光强度明显降低.结论:CHE能够抑制人卵巢癌SKOV3 细胞增殖、迁移、侵袭和EMT.

Abstract

Objective:To discuss the inhibitory effect of chelerythrine(CHE)on the migration,invasion,and epithelial-mesenchymal transition(EMT)of the human ovarian cancer SKOV3 cells,and to clarify the associated mechanism.Methods:The SKOV3 cells were cultured in vitro and divided into control group and 2.5,5.0,10.0,20.0,and 40.0 μmol·L-1 CHE groups.Methylthiazolydiphenyl-tetrazolium(MTT)assay was used to detect the inhibitory rates of proliferation of the cells in various groups.The SKOV3 cells were cultured in vitro and divided into control group,transforming growth factor-β1(TGF-β1)group,TGF-β1+5 μmol·L-1 CHE group,and TGF-β1+10 μmol·L-1 CHE group.Cell scratch assay was used to detect the migration rates of the cells in various groups;Transwell chamber assay was used to detect the numbers of migration and invasion cells in various groups;Western blotting method was used to detect the expression levels of E-cadherin,N-cadherin,and Vimentin proteins in the cells in various groups;immunofluorescence staining method was used to detect the fluorescence intensities of E-cadherin and N-cadherin in the cells in various groups.Results:The MTT assay results showed that compared with control group,the inhibitory rates of proliferation of the cells in 5.0,10.0,20.0,and 40.0 μmol·L-1 CHE groups were significantly increased(P<0.05 or P<0.01).The cell scratch assay results showed that compared with control group,the migration rate of the cells in TGF-β1 group was increased(P<0.01);compared with TGF-β1 group,the migration rates of the cells in TGF-β1+5 μmol·L-1 CHE group and TGF-β1+10 μmol·L-1 CHE group were significantly decreased(P<0.01).The Transwell chamber assay results showed that compared with control group,the numbers of migration and invasion cells in TGF-β1 group were significantly increased(P<0.05);compared with TGF-β1 group,the numbers of migration and invasion cells in TGF-β1+5 μmo·l L-1 CHE group and TGF-β1+10 μmo·l L-1 CHE group were significantly decreased(P<0.01).The Western blotting results showed that compared with control group,the expression level of E-cadherin protein in the cells in TGF-β1 group was significantly decreased(P<0.01),while the expression levels of N-cadherin and Vimentin proteins were increased(P<0.05 or P<0.01);compared with TGF-β1 group,the expression levels of E-cadherin protein in the cells in TGF-β1+5 μmol·L-1 CHE group and TGF-β1+10 μmol·L-1 CHE group were significantly increased(P<0.01),and the expression levels of N-cadherin and Vimentin proteins were significantly decreased(P<0.01).The immunofluorescence staining results showed that compared with control group,the fluorescence intensity of E-cadherin in the cells in TGF-β1 group was decreased,and the fluorescence intensity of N-cadherin was increased;compared with TGF-β1 group,the fluorescence intensities of E-cadherin in the cells in TGF-β 1+5 μmol·L-1 CHE group and TGF-β1+10 μmol·L-1 CHE group were significantly increased,and the fluorescence intensities of N-cadherin were decreased.Conclusion:CHE can inhibit the proliferation,migration,invasion,and EMT of the human ovarian cancer SKOV3 cells.

关键词

白屈菜红碱/卵巢肿瘤/上皮-间质转化/转化生长因子β1/细胞迁移/细胞侵袭

Key words

Chelerythrine/Ovarian neoplasm/Epithelial-mesenchymal transition/Transforming growth factor-β1/Cell migration/Cell invasion

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基金项目

吉林省科技厅科技发展计划项目(20210402034GH)

吉林省发改委创新能力建设项目(2021C011)

出版年

2024
吉林大学学报(医学版)
吉林大学

吉林大学学报(医学版)

CSTPCDCSCD北大核心
影响因子:0.909
ISSN:1671-587X
参考文献量4
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