沉默E26转录因子变异体4抑制核因子-κB信号通路对结肠癌细胞增殖和迁移的影响

徐和希; 宋红旗; 刘佃温; 刘世举; 杨会举

doi:10.7619/jcmp.20243267

沉默E26转录因子变异体4抑制核因子-κB信号通路对结肠癌细胞增殖和迁移的影响

1.
河南中医药大学, 河南郑州, 450046
2.
河南中医药大学第三附属医院肛肠科, 河南郑州, 450046

基金项目:

河南省中医药拔尖人才项目豫卫中医函[2021]15号

河南省中医药科学研究专项课题 2023ZY2116

详细信息

通讯作者:
杨会举

中图分类号: R735.3;R730.43;Q344
计量
- 文章访问数: 61
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出版历程
- 收稿日期: 2024-07-30
- 修回日期: 2024-10-27
- 刊出日期: 2025-01-27

Effect of silencing E26 transformation-specific sequence 4 on proliferation and migration of colon cancer cells by inhibiting nuclear factor-κB signaling pathway

1.
Henan University of Traditional Chinese Medicine, Zhengzhou, Henan, 450046
2.
Anorectal Department, the Third Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou, Henan, 450046

摘要

摘要:
目的
探讨E26转录因子变异体4(ETV4)通过核因子-κB(NF-κB)信号通路对结肠癌细胞增殖和迁移的影响机制。
方法
通过用户友好的交互式癌症转录组数据分析资源(UALCAN)数据库分析ETV4在结肠正常组织和癌组织中的表达; 采用逆转录定量聚合酶链式反应(qRT-PCR)和Western blot检测ETV4在正常肠上皮细胞和结肠癌细胞系中的表达; 沉默SW480细胞的ETV4后, 采用qRT-PCR和Western blot检测ETV4的表达以评估转染效率; 采用菌落形成实验和Transwell实验检测沉默ETV4后对结肠癌细胞增殖和迁移的影响; 采用Western blot检测沉默ETV4后对NF-κB通路中的蛋白65(p65)和磷酸化蛋白65(p-p65)的蛋白表达的影响。
结果
UALCAN数据库分析结果显示ETV4在结肠癌组织中高表达。qRT-PCR和Western blot检测显示ETV4在结肠癌细胞系SW480、Lovo、Caco-2和SW620中的表达高于正常肠上皮细胞HIEC-6, 其中SW480细胞中ETV4的表达最高，差异有统计学意义(P < 0.001)。菌落形成实验和Transwell实验结果显示，沉默ETV4后显著抑制了结肠癌细胞SW480的增殖和迁移能力(P < 0.001)。Western blot检测结果显示，沉默ETV4显著抑制细胞中p-p65蛋白的表达(P < 0.001)。
结论
沉默ETV4可能抑制NF-κB信号通路的激活，进而抑制结肠癌细胞的增殖和迁移。
- E26转录因子变异体4 /
- 核因子-κB /
- 结肠癌 /
- 细胞增殖 /
- 细胞迁移
Abstract:
Objective
To investigate the mechanism of E26 transformation-specific sequence 4 (ETV4) affecting the proliferation and migration of colon cancer cells through the nuclear factor-κB (NF-κB) signaling pathway.
Methods
The expression level of ETV4 in normal colon tissues and cancer tissues was analyzed by the user-friendly interactive cancer transcriptome data analysis resource (UALCAN) database. Reverse transcription quantitative polymerase chain reaction (qRT-PCR) and Western blot were used to detect the expression level of ETV4 in normal intestinal epithelial cells and colon cancer cell lines. After silencing ETV4 in SW480 cells, qRT-PCR and Western blot were performed to detect the expression of ETV4 to assess transfection efficiency; colony formation and Transwell assays were conducted to explore the effects of ETV4 silencing on the proliferation and migration of colon cancer cells; the Western blot was used to detect the effects of ETV4 silencing on the protein expression of protein 65 (p65) and phosphorylated protein 65 (p-p65) in the NF-κB pathway.
Results
The UALCAN database analysis revealed high expression of ETV4 in colon cancer tissues. The qRT-PCR and Western blot showed that ETV4 expression was significantly higher in the colon cancer cell lines SW480, Lovo, Caco-2, and SW620 than in normal intestinal epithelial cells HIEC-6, with the highest expression in SW480 cells (P < 0.001). Colony formation and Transwell assay results indicated that silencing ETV4 significantly inhibited the proliferation and migration of colon cancer SW480 cells (P < 0.001). Western blot results showed that silencing ETV4 significantly inhibited the expression of p-p65 protein in the cells (P < 0.001).
Conclusion
Silencing ETV4 may inhibit the activation of the NF-κB signaling pathway, thereby inhibiting the proliferation and migration of colon cancer cells.
- E26 transformation-specific sequence 4 /
- nuclear factor-κB /
- colon cancer /
- cell proliferation /
- cell migration

HTML全文

图 1 ETV4在结肠癌中的表达

A: ETV4在结肠癌组织与正常结肠组织中的表达; B: ETV4在4种结肠癌细胞和正常肠上皮细胞中的表达。

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图 2 sh-ETV4组与sh-NC组ETV4的蛋白表达

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图 3 sh-NC组与sh-ETV4组菌落形成实验结果

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图 4 sh-NC组与sh-ETV4组Transwell实验结果

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图 5 Western blot实验检测NF-κB通路蛋白表达

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表 1 各细胞中 ETV4 mRNA和ETV4蛋白相对表达(x±s)

细胞	ETV4 mRNA相对表达	ETV4蛋白相对表达
HIEC-6	1.00±0.15	0.26±0.03
SW480	3.22±0.36^***	0.95±0.08^***
Lovo	2.64±0.19^***	0.59±0.07^***
Caco-2	1.67±0.15^*	0.42±0.04^*
SW620	2.44±0.14^***	0.51±0.06^**
ETV4: E26转录因子变异体4。与HIEC-6比较, * P < 0.05, * * P < 0.01, * * * P < 0.001

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表 2 sh-NC组与sh-ETV4组 ETV4 mRNA和ETV4蛋白表达比较(x±s)

组别	ETV4 mRNA相对表达	ETV4蛋白相对表达
sh-NC组	1.00±0.11	0.84±0.07
sh-ETV4组	0.38±0.04^***	0.28±0.04^***
与sh-NC组比较, * * * P < 0.001。

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