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| MiR-579-3p/ MYLK通路在胃癌细胞凋亡中的作用及机制 |
| The role and mechanism of miR-579-3p/ MYLK pathway in the apoptosis of gastric cancer cells |
| 投稿时间：2025-03-25 修订日期：2025-07-08 |
| DOI： |
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| 中文关键词: 胃癌 微小RNA-579-3p 凋亡 双荧光素酶报告基因分析 生物信息学 |
| 英文关键词:Gastric carcinoma miR-579-3p apoptosis luciferase reporter assay bioinformatics. |
| 基金项目:河北省省级科技计划资助(22377701D)、河北省医学科学研究课题计划资助（20210886） |
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| 摘要点击次数: 235 |
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| 中文摘要: |
| 目的 探讨MiR-579-3p（微小RNA-579-3p，MicroRNA-579-3p）及其靶基因MYLK（肌球蛋白轻链激酶，myosin light chain kinase）在胃癌细胞凋亡调控中的作用及相关分子机制。方法 生物信息学分析MiR-579-3p的表达及功能。实时定量PCR检测MiR-579-3p及相关基因表达水平。应用MiR-579-3p抑制物转染人胃癌细胞株HGC-27；CCK-8法检测细胞活性；流式细胞术检测细胞凋亡率；蛋白印迹法检测凋亡相关基因及蛋白的表达。双荧光素酶报告基因分析验证MiR-579-3p直接调控的基因。结果 胃癌组织中MiR-579-3p水平明显高于癌旁正常组织，验证结果与生信结果符合（P<0.05）。胃癌细胞株的MiR-579-3p表达均高于正常胃上皮细胞株GES-1，其中HGC-27细胞MiR-579-3p表达水平最高（P<0.05）。通过对MiR-579-3p的功能进行富集分析，发现MiR-579-3p与多种生物学过程及通路有关；MYLK在肿瘤组织表达低于癌旁组织(P<0.001)，MYLK表达与MiR-579-3p负相关；胃癌细胞株除细胞株MKN45外，MYLK水平均低于GES-1(P<0.01)。在HGC-27细胞中抑制MiR-579-3p表达后MYLK表达增高（P<0.05）。双荧光素酶报告基因分析结果发现MYLK为MiR-579-3p直接调控的基因。通过CCK-8和流式细胞术检测验证抑制MiR-579-3p表达可抑制细胞增殖（P<0.001）和促进癌细胞凋亡（P<0.05）。转染MiR-579-3p抑制物后HGC-27细胞中Bcl-2（P<0.05）、Survivin（P<0.05）的mRNA和蛋白表达明显降低，而Bax的表达明显增高（P<0.001）。结论 MiR-579-3p能通过抑制MYLK表达，进而调节Bcl-2、Survivin、Bax等凋亡基因而参与胃癌细胞的凋亡过程。 |
| 英文摘要: |
| Objective: To investigate the role and molecular mechanisms of MiR-579-3p (MicroRNA-579-3p) and its target gene MYLK (myosin light chain kinase) in the regulation of apoptosis in gastric cancer cells. Methods: Bioinformatics analysis was conducted to assess the expression and function of MiR-579-3p. Quantitative real-time PCR was employed to measure the expression levels of MiR-579-3p and related genes. The human gastric cancer cell line HGC-27 was transfected with a MiR-579-3p inhibitor. Cell viability was assessed using the CCK-8 assay, and apoptosis rates were determined by flow cytometry. Western blotting was used to detect the expression of apoptosis-related genes and proteins. Dual-luciferase reporter gene analysis was performed to validate the genes directly regulated by MiR-579-3p. Results: The level of MiR-579-3p in gastric cancer tissues was significantly higher than in adjacent normal tissues, consistent with bioinformatics results (P<0.05). The expression of MiR-579-3p in gastric cancer cell lines was higher than in the normal gastric epithelial cell line GES-1, with the highest expression observed in HGC-27 cells (P<0.05). Functional enrichment analysis revealed that MiR-579-3p is associated with various biological processes and pathways. MYLK expression was lower in tumor tissues compared to adjacent tissues (P<0.001) and negatively correlated with MiR-579-3p. MYLK levels in gastric cancer cell lines, except for MKN45, were lower than in GES-1 (P<0.01). Inhibition of MiR-579-3p expression in HGC-27 cells led to increased MYLK expression (P<0.05). Dual-luciferase reporter gene analysis confirmed MYLK as a direct target of MiR-579-3p. CCK-8 and flow cytometry assays demonstrated that inhibiting MiR-579-3p expression suppressed cell proliferation (P<0.001) and promoted cancer cell apoptosis (P<0.05). Transfection with the MiR-579-3p inhibitor significantly reduced the mRNA and protein expression of Bcl-2 (P<0.05) and Survivin (P<0.05) while increasing Bax expression (P<0.001) in HGC-27 cells. Conclusion: MiR-579-3p participates in the apoptosis process of gastric cancer cells by inhibiting MYLK expression and subsequently regulating apoptosis-related genes such as Bcl-2, Survivin, and Bax. |
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