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胡忠源, 张成明, 鱼涛.雷公藤红素通过miR-224-5p/EGR2通路抑制胃癌MGC803细胞生物学行为的作用机制[J].湖南中医药大学学报,2025,45(11):2081-2089[点击复制] |
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| 雷公藤红素通过miR-224-5p/EGR2通路抑制胃癌MGC803细胞生物学行为的作用机制 |
| 胡忠源,张成明,鱼涛 |
| (陕西省中医医院脾胃一科, 陕西 西安 710000) |
| 摘要: |
| 目的 探讨雷公藤红素（CL）对胃癌MGC803细胞增殖、凋亡和侵袭的影响及其机制。方法 体外培养人胃癌MGC803细胞，设立对照组、CL(0.5、1.0、2.0、4.0μmol/L)组、微小RNA-224-5p(mi R-224-5p)抑制组（mi R-224-5p-i组）、CL联合mi R-224-5p过表达组（CL+mimic组）、CL联合si-EGR2组（CL+si-EGR2组）及mi R-224-5p抑制联合si-EGR2组（mi R-224-5p-i+si-EGR2组）。通过CCK-8法、流式细胞术、Transwell侵袭实验及划痕愈合实验评估细胞增殖、凋亡及迁移侵袭能力；RT-qPCR及Western blot检测mi R-224-5p、早期生长反应因子2(EGR2)及其下游蛋白细胞周期蛋白D1(Cyclin D1)、切割型胱天蛋白酶-3(cleaved Caspase-3)、基质金属蛋白酶-9(MMP-9)表达水平；双荧光素酶报告实验验证mi R-224-5p对EGR2的靶向调控关系。结果 与对照组相比，CL处理后MGC803细胞活力、迁移细胞数、侵袭细胞数、划痕愈合率及Cyclin D1、MMP-9蛋白表达水平降低（P<0.01），凋亡率及EGR2、cleaved Caspase-3蛋白表达水平升高（P<0.01）;mi R-224-5p表达下调（P<0.01）。双荧光素酶实验结果证实，mi R-224-5p可特异性结合EGR2 3'UTR区调控其表达。与CL 2.0μmol/L组比较，CL+mimic组细胞活力、划痕愈合率及Cyclin D1、MMP-9蛋白表达升高（P<0.05），凋亡率及EGR2、cleaved Caspase-3蛋白表达降低（P<0.05）。与mi R-224-5p-i组比较，mi R-224-5p-i+si-EGR2组细胞活力、划痕愈合率及Cyclin D1、MMP-9表达升高（P<0.05），凋亡率及EGR2、cleaved Caspase-3表达降低（P<0.05）。结论 CL可通过下调miR-224-5p表达，解除其对EGR2的抑制，进而抑制胃癌MGC803细胞增殖与侵袭，诱导凋亡。 |
| 关键词: 胃癌 微小RNA-224-5p 早期生长反应因子2 增殖 凋亡 侵袭 |
| DOI：10.3969/j.issn.1674-070X.2025.11.008 |
| 投稿时间：2025-05-08 |
| 基金项目:陕西省重点研发计划项目(2024SF-YBXM-499)。 |
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| Mechanism of action of celastrol inhibiting biological behaviors of gastric cancer MGC803 cells via the miR-224-5p/EGR2 pathway |
| HU Zhongyuan, ZHANG Chengming, YU Tao |
| (The First Department of Spleen and Stomach Diseases, Shaanxi Provincial Hospital of Chinese Medicine, Xi'an, Shaanxi 710000, China) |
| Abstract: |
| Objective To investigate the effects of celastrol（CL） on the proliferation, apoptosis, and invasion of gastric cancer MGC803 cells and its underlying mechanism. Methods Human gastric cancer MGC803 cells were cultured in vitro and divided into control group, CL（0.5, 1.0, 2.0, 4.0 μmol/L） groups, mi R-224-5p inhibitor group（mi R-224-5p-i group）, CL combined with miR-224-5p overexpression group（CL+mimic group）, CL combined with si-EGR2 group（CL+si-EGR2 group）, and miR-224-5p inhibitor combined with si-EGR2 group（miR-224-5p-i+si-EGR2 group）. Cell proliferation, apoptosis, migration and invasion capacities were assessed using the CCK-8 assay, flow cytometry, Trans well invasion assay, and scratch wound healing assay respectively. The expression levels of miR-224-5p, early growth response 2（EGR2）, and its downstream proteins Cyclin D1, Cleaved Caspase-3, and matrix metalloproteinase-9（MMP-9） were measured by RT-q PCR and Western blot. The targeting relationship between mi R-224-5p and EGR2 was verified using a dual-luciferase reporter assay. Results Compared with the control group, CL treatment resulted in reduced cell viability, number of migrating and invading cells, scratch wound healing rate, and the protein expression levels of Cyclin D1 and MMP-9 in MGC803 cells（P＜0.01）, while it increased the apoptosis rate and the protein expression levels of EGR2 and Cleaved Caspase-3（P＜0.01）, and decreased the expression level of mi R-224-5p（P＜0.01）. The dual-luciferase reporter assay confirmed that miR-224-5p specifically binds to the 3'UTR region of EGR2 to regulate its expression. Compared with the CL 2.0 μmol/L group, the CL+mimic group exhibited increased cell viability, scratch wound healing rate, and protein expression levels of Cyclin D1 and MMP-9（P＜0.05）, while the apoptosis rate and protein expression levels of EGR2 and Cleaved Caspase-3 decreased（P＜0.05）.Compared with the miR-224-5p-i group, the miR-224-5p-i+si-EGR2 group showed elevated cell viability, scratch wound healing rate, and expression levels of Cyclin D1 and MMP-9（P＜0.05）, along with reduced apoptosis rate and expression levels of EGR2 and Cleaved Caspase-3（P＜0.05）. Conclusion CL can inhibit the proliferation and invasion of MGC803 gastric cancer cells and induce apoptosis by downregulating miR-224-5p expression and relieving its inhibitory effects on EGR2. |
| Key words: gastric cancer miRNA-224-5p early growth response factor 2 proliferation apoptosis invasion |
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