草苁蓉环烯醚萜苷（IGBR）对TGF-β1诱导的HepG2细胞上皮间质转化模型的影响

金爱花; 朱洁波; 尹学哲; 全吉淑

doi:10.12449/JCH240617

草苁蓉环烯醚萜苷（IGBR）对TGF-β1诱导的HepG2细胞上皮间质转化模型的影响

DOI: 10.12449/JCH240617

金爱花^1a,
朱洁波²,
尹学哲^1b,
全吉淑^2, , ,

1a.
延边大学附属医院（延边医院）医学检验科延吉吉林 133002
1b.
延边大学附属医院（延边医院）呼吸与危重症医学科延吉吉林 133002
2.
延边大学医学院生物化学与分子生物学教研室延吉吉林 133002

基金项目:

国家自然科学基金 (82060113);

国家自然科学基金 (81760659);

吉林省教育厅项目 (JJKH20210578KJ)

利益冲突声明：本文不存在任何利益冲突。

作者贡献声明：金爱花负责课题设计，资料分析，撰写论文，参与实验；朱洁波参与数据分析与实验；尹学哲参与数据分析与修改论文；全吉淑负责拟定写作思路，指导撰写文章并最后定稿。

详细信息

通信作者:
全吉淑， quanjs@ybu.edu.cn （ORCID： 0000-0002-6466-5787）

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出版历程
- 收稿日期: 2024-01-23
- 录用日期: 2024-02-28
- 出版日期: 2024-06-25

Effect of iridoid glycosides from Boschniakia rossica on epithelial-mesenchymal transition of HepG2 cells induced by transforming growth factor-beta 1

Aihua JIN^1a,
Jiebo ZHU²,
Xuezhe YIN^1b,
Jishu QUAN^{2
, ,
,}

1a.
Department of Clinical Laboratory,The Affiliated Hospital of Yanbian University （Yanbian Hospital）, Yanji, Jilin 133002
1b.
Department of Respiratory and Critical Care Medicine,The Affiliated Hospital of Yanbian University （Yanbian Hospital）, Yanji, Jilin 133002
2.
Department of Biochemistry and Molecular Biology，Yanbian University Medical College，Yanji, Jilin 133002，China

Research funding:

National Natural Science Foundation of China (82060113);

National Natural Science Foundation of China (81760659);

Jilin Province Education Department project (JJKH20210578KJ)

More Information

Corresponding author: QUAN Jishu， quanjs@ybu.edu.cn （ORCID： 0000-0002-6466-5787）

摘要

摘要: 目的研究草苁蓉环烯醚萜苷（IGBR）对TGF-β1诱导肝癌HepG2细胞上皮间质转化（EMT）的影响作用。方法用10 μg/L TGF-β1诱导HepG2肝癌细胞株构建肝癌细胞EMT模型。实验分为对照组、模型组与IGBR组3组，对照组用无血清DMEM处理，模型组用10 μg/L TGF-β1处理，IGBR组用10 μg/L TGF-β1和500 mg/L IGBR联合处理，培养48 h。利用细胞黏附实验、划痕愈合实验和Transwell小室实验观察细胞迁移和侵袭能力。RT-PCR法和Western Blot法检测细胞中E-钙黏蛋白、N-钙黏蛋白、波形蛋白的mRNA和蛋白表达，Western Blot法检测Slug、Twist1、ZEB1、p-STAT3、STAT3的蛋白表达。计量资料多组间比较采用单因素方差分析，进一步两两比较采用LSD-t检验；两组间比较采用成组t检验。结果 TGF-β1诱导后，模型组HepG2细胞呈现长梭形改变；与模型组比较，IGBR组细胞黏附率降低，抑制细胞迁移、侵袭能力（P值均<0.05），E-钙黏蛋白的mRNA表达和蛋白表达均增高（P值均<0.05），N-钙黏蛋白和波形蛋白的mRNA表达和蛋白表达均降低（P值均<0.05），Slug、Twist1、ZEB1蛋白表达和p-STAT3蛋白表达均降低（P值均<0.05）。结论 IGBR可抑制TGF-β1诱导的HepG2细胞EMT过程，从而减弱HepG2细胞黏附力和细胞迁移、侵袭能力，上调E-钙黏蛋白，下调N-钙黏蛋白和波形蛋白，上调Slug、Twist1、ZEB1、STAT3的蛋白表达，其作用可能通过抑制STAT3通路下调Slug、Twist1、ZEB1等EMT转录因子来实现。
- 肝肿瘤 /
- 列当 /
- 环烯醚萜类 /
- 上皮-间质转化
Abstract: Objective To investigate the effect of iridoid glycosides from Boschniakia rossica （IGBR） on epithelial-mesenchymal transition （EMT） of HepG2 hepatoma cells induced by transforming growth factor-beta 1 （TGF-β1）. Methods HepG2 hepatoma cells were induced by 10 μg/L TGF-β1 to construct an EMT model of hepatoma cells. The cells were divided into control group （treated with serum-free DMEM）， model group （treated with 10 μg/L TGF-β1）， and IGBR group （treated with 10 μg/L TGF-β1 and 500 mg/L IGBR）， and all cells were cultured for 48 hours. Cell adhesion assay， wound healing assay， and Transwell chamber assay were used to observe the migration and invasion abilities of cells. RT-PCR and Western blot were used to measure the mRNA and protein expression levels of E-cadherin， N-cadherin， and vimentin in cells， and Western blot was used to measure the protein expression levels of Slug， Twist1， ZEB1， p-STAT3， and STAT3. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups； the independent-samples t test was used for comparison between two groups. Results After TGF-β1 induction， HepG2 cells in the model group showed long spindle-shape changes， while those in the control group showed polygonal epithelia-like changes. Compared with the model group， the IGBR group had a significant reduction in cell adhesion rate and significant inhibition of cell migration and invasion abilities （all P<0.05）， as well as significant increases in the mRNA and protein expression levels of E-cadherin （P<0.05）， significant reductions in the mRNA and protein expression levels of N-cadherin and vimentin （all P<0.05）， and significant reductions in the protein expression levels of Slug， Twist1， ZEB1， and p-STAT3 （all P<0.05）. Conclusion IGBR can inhibit TGF-β1-induced EMT process in HepG2 cells， thereby attenuating cell adhesion， migration， and invasion abilities， and it can also upregulate E-cadherin， downregulate N-cadherin and vimentin， and upregulate the protein expression of Slug， Twist1， ZEB1， and STAT3， possibly by inhibiting the STAT3 pathway to downregulate the EMT transcription factors such as Slug， Twist1， and ZEB1.
- Liver Neoplasms /
- Orobanche Coerulescens /
- Iridoids /
- Epithelial-Mesenchymal Transition

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Figure . The EMT model of HepG2 cells was induced by TGF-β1

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图 2 IGBR对HepG2细胞形态的影响（×100）

注： a，对照组；b，模型组；c，IGBR组。

Figure 2. Effect of IGBR on HepG 2 cell morphology（×100）

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图 3 IGBR对HepG2细胞迁移能力的影响（×40）

Figure 3. Effect of IGBR on the migratory ability of HepG 2 cells（×40）

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图 4 IGBR对HepG2细胞侵袭能力的影响（×100）

注： a，对照组；b，模型组；c，IGBR组。

Figure 4. Effect of IGBR on the invasive ability of HepG 2 cells（×100）

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图 5 IGBR对HepG2细胞E-钙黏蛋白、N-钙黏蛋白和波形蛋白表达的影响

注： a，RT-PCR结果；b，Western Blot结果。

Figure 5. Effect of IGBR on the expression of E-cadherin， N-cadherin， and vimentin in HepG 2 cells

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图 6 IGBR对HepG2细胞EMT相关转录因子表达的影响

Figure 6. Effect of IGBR on the expression of EMT-associated transcription factors in HepG 2 cells

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图 7 IGBR对HepG2细胞STAT3蛋白表达的影响

Figure 7. Effect of IGBR on STAT 3 protein expression in HepG 2 cells

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表 1 HepG2细胞EMT模型中E-钙黏蛋白、N-钙黏蛋白和波形蛋白表达的比较

Table 1. Comparison of E-cadherin， N-cadherin， and vimentin expression in the EMT model of HepG2 cells

组别	mRNA			蛋白
组别	E-钙黏蛋白	N-钙黏蛋白	波形蛋白	E-钙黏蛋白	N-钙黏蛋白	波形蛋白
对照组	1.41±0.11	0.25±0.06	0.36±0.08	1.40±0.13	0.42±0.06	0.36±0.12
模型组	0.26±0.09	1.45±0.14	0.89±0.10	0.31±0.09	1.63±0.14	2.10±0.10
t值	14.015	13.646	7.168	11.940	13.759	19.294
P值	0.000 2	0.000 2	0.002 0	0.000 3	0.000 2	<0.000 1

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表 2 IGBR对HepG2细胞E-钙黏蛋白、N-钙黏蛋白和波形蛋白表达的比较

Table 2. Comparison of IGBR on E-cadherin， N-cadheractin， and vimentin expression in HepG 2 cells

组别	mRNA			蛋白
组别	E-钙黏蛋白	N-钙黏蛋白	波形蛋白	E-钙黏蛋白	N-钙黏蛋白	波形蛋白
对照组	0.80±0.05	0.17±0.06	0.14±0.04	1.45±0.05	0.12±0.06	0.14±0.05
模型组	0.20±0.09^1）	0.78±0.06^1）	1.20±0.10^1）	0.25±0.03^1）	1.08±0.09^1）	1.37±0.10^1）
IGBR组	0.42±0.07^2）	0.58±0.03^2）	0.65±0.09^2）	0.08±0.10^2）	0.70±0.12^2）	1.10±0.14^2）
F值	53.50	107.40	128.40	242.40	69.01	127.90
P值	0.000 1	<0.000 1	<0.000 1	<0.000 1	<0.000 1	<0.000 1
注：与对照组相比，1） P<0.05；与模型组相比，2） P<0.05。

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