过表达lncRNA HCG11调控线粒体自噬抑制喉癌细胞恶性生物学行为的机制研究

Study on the mechanism by which upregulation of lncRNA HCG11 regulates mitochondrial autophagy and suppresses malignant biological behaviors in laryngeal cancer cells

摘要:

目的 探讨长链非编码RNA HLA复合物11组(lncRNA HCG11)对HEp-2喉癌细胞恶性生物学行为的影响及机制。方法 利用质粒转染技术将人喉表皮样癌HEp-2细胞分为lncRNA HCG11对照组和过表达组,人鼻咽上皮细胞NP69细胞作为正常组。采用qPCR检测HCG11在三组细胞中表达情况。通过克隆形成实验检测细胞增殖；划痕实验检测细胞迁移；Annexin V/PI染色检测细胞凋亡；DCFH-DA荧光探针染色检测细胞内活性氧(ROS)；透射电镜检测细胞内线粒体；Western blot分析细胞中PINK1、Parkin、LC3Ⅱ/LC3Ⅰ、p62蛋白表达。结果 qPCR结果显示,喉癌细胞比正常细胞HCG11 mRNA水平表达更低,且成功构建喉癌细胞HCG11 mRNA过表达模型。与对照组比,HCG11过表达组HEp-2细胞增殖、迁移及凋亡均被抑制(P<0.05)；细胞内ROS水平升高(P<0.05)；细胞内线粒体形态受损；细胞PINK1、Parkin、LC3Ⅱ/LC3Ⅰ蛋白水平升高,p62蛋白水平降低(P<0.05)。结论 LncRNA HCG11在HEp-2喉癌细胞中表达下调,lncRNA HCG11过表达可抑制HEp-2喉癌细胞增殖、迁移及凋亡,其机制可能与激活PINK1/Parkin信号通道,从而调控胞内线粒体发生过度自噬相关。

Abstract:

Objective Exploring the effect and mechanism of lncRNA HCG11 on the malignant biological behavior of HEp-2 laryngeal cancer cells. Methods Through plasmid transfection, HEp-2 cells from human laryngeal squamous cell carcinoma were separated into a control and an HCG11 overexpression group, with NP69 nasopharyngeal epithelial cells serving as the normal control. Expression levels of HCG11 in these cell groups were assessed by QPCR. Cellular proliferation was measured via cloning formation assays, migration was monitored using scratch tests, apoptosis was identified through Annexin V/PI staining, and intracellular ROS levels were quantified with the DCFH-DA fluorescent probe. Mitochondrial structure was visualized under transmission electron microscopy, and the protein expression of PINK1, Parkin, the LC3 II/I ratio, and p62 was determined by Western blot. Results