目的 探究网状细胞受体4（reticulon 4,RTN4）对HeLa细胞增殖、凋亡以及迁移的影响,为宫颈癌发生发展过程中的分子基础和治疗靶点提供新的理论依据。方法 通过TCGA（the cancer genome atlas）、GTEx（genotype-tissue expression）、GEO（gene expression omnibus）、HPA（human protein atlas）等数据库以及免疫组织化学染色方法,鉴定宫颈癌和癌旁组织中RTN4 mRNA及蛋白的表达差异;HeLa细胞中上调及下调RTN4水平后,通过qPCR、Western Blotting、CCK-8试验、克隆形成试验、划痕试验、侵袭试验、细胞凋亡试验等,观察RTN4对HeLa细胞增殖、凋亡及迁移的影响。结果 数据库分析发现,与癌旁组织相比,宫颈癌组织中RTN4 mRNA和蛋白水平显著下降;免疫组织化学染色结果显示,相对于癌旁组织,RTN4蛋白在宫颈癌中显著低表达;在HeLa细胞中,上调RTN4水平后,细胞的增殖、迁移、侵袭能力显著降低,凋亡率显著升高;而下调RTN4水平后,细胞的增殖、迁移、侵袭能力显著升高,凋亡率则显著降低。结论 在宫颈癌组织中,RTN4 mRNA和蛋白表达显著低于癌旁组织。在宫颈癌细胞中,RTN4过表达可抑制细胞的增殖、迁移、侵袭,并促进其凋亡;而下调RTN4则呈现相反表型。这些结果提示RTN4可能成为宫颈癌靶向治疗的新靶点。
Abstract
Objective To investigate the effects of RTN4 on the proliferation, apoptosis, and migration of HeLa cells, providing a new theoretical basis for the molecular mechanisms and therapeutic targets in the development of cervical cancer. Methods The expression differences of RTN4 mRNA and protein in cervical cancer and adjacent tissues were identified through databases such as TCGA, GTEx, GEO, and HPA, as well as immunohistochemical staining. After upregulating and downregulating RTN4 levels in HeLa cells, the effects of RTN4 on cell proliferation, apoptosis, and migration were observed using qPCR, Western Blotting, CCK-8 assay, colony formation assay, scratch assay, invasion assay, and apoptosis assays. Results Database analysis revealed that the levels of RTN4 mRNA and protein were significantly decreased in cervical cancer tissues compared to adjacent tissues. Immunohistochemical staining results showed that RTN4 protein was significantly underexpressed in cervical cancer compared to adjacent tissues. In HeLa cells, upregulating RTN4 levels led to a significant decrease in cell proliferation, migration, and invasion abilities, along with a significant increase in apoptosis rate. Conversely, downregulating RTN4 levels resulted in a significant increase in cell proliferation, migration, and invasion abilities, while the apoptosis rate significantly decreased. Conclusion The expression of RTN4 mRNA and protein in cervical cancer tissues is significantly lower than that in adjacent tissues. In cervical cancer cells, overexpression of RTN4 can inhibit cell proliferation, migration, and invasion, while promoting apoptosis; downregulation of RTN4 exhibits the opposite phenotype. These results suggest that RTN4 may serve as a new target for targeted therapy in cervical cancer.
关键词
宫颈癌 /
网状细胞受体4 /
增殖 /
迁移 /
凋亡
Key words
cervical cancer /
RTN4 /
proliferation /
migration /
apoptosis
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基金
湖南省中医药管理局基金项目“基于代谢组学的孕妇中医体质与婴儿湿疹的相关性研究”（C2023046）
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