Department of Pediatric Surgery, Henan Provincial People's Hospital, Zhengzhou 450000, China.
Department of Obstetrics, Henan Provincial People's Hospital, Zhengzhou 450000, China.
Anal Cell Pathol (Amst). 2023 Jul 1;2023:6761894. doi: 10.1155/2023/6761894. eCollection 2023.
To study the effect of congenital dyskeratosis 1 (DKC1) on neuroblastoma and its regulation mechanism.
The expression of DKC1 in neuroblastoma was analyzed by TCGA database and molecular assay. NB cells were transfected with siDKC1 to observe the effects of DKC1 on proliferation, cloning, metastasis, and invasion, and apoptosis and apoptosis-related proteins. The tumor-bearing mouse model was constructed, shDKC1 was transfected to observe the tumor growth and tumor tissue changes, and the expression of DKC1 and Ki-67 was detected. Screening and identification of miRNA326-5p targeting DKC1. NB cells were treated with miRNA326-5p mimic or inhibitors to detect the expression of DKC1. NB cells were transfected with miRNA326-5p and DKC1 mimics to detect cell proliferation, apoptosis, and apoptotic protein expression.
DKC1 was highly expressed in NB cells and tissues. The activity, proliferation, invasion, and migration of NB cells were significantly decreased by DKC1 gene knockout, while apoptosis was significantly increased. The expression level of B-cell lymphoma-2 in shDKC1 group was significantly lower than that of the control group, while the expression level of BAK, BAX, and caspase-3 was significantly higher than that of the control group. The results of experiments on tumor-bearing mice were consistent with the above results. The results of miRNA assay showed that miRNA326-5p could bind DKC1 mRNA to inhibit the protein expression, thereby inhibiting the proliferation of NB cells, promoting their apoptosis, and regulating the expression of apoptotic proteins.
miRNA326-5p targeting DKC1 mRNA regulates apoptosis-related proteins to inhibit neuroblastoma proliferation and promote the apoptotic process.
研究先天性角化不良 1(DKC1)对神经母细胞瘤的影响及其调控机制。
通过 TCGA 数据库和分子检测分析神经母细胞瘤中 DKC1 的表达。用 siDKC1 转染 NB 细胞,观察 DKC1 对增殖、克隆形成、转移、侵袭和凋亡及凋亡相关蛋白的影响。构建荷瘤小鼠模型,转染 shDKC1 观察肿瘤生长和肿瘤组织变化,并检测 DKC1 和 Ki-67 的表达。筛选和鉴定靶向 DKC1 的 miRNA326-5p。用 miRNA326-5p 模拟物或抑制剂处理 NB 细胞,检测 DKC1 的表达。用 miRNA326-5p 和 DKC1 模拟物转染 NB 细胞,检测细胞增殖、凋亡和凋亡蛋白表达。
DKC1 在 NB 细胞和组织中高表达。DKC1 基因敲除后,NB 细胞的活性、增殖、侵袭和迁移能力明显下降,凋亡明显增加。shDKC1 组的 B 细胞淋巴瘤-2 表达水平明显低于对照组,而 BAK、BAX 和 caspase-3 的表达水平明显高于对照组。荷瘤小鼠实验结果与上述结果一致。miRNA 检测结果表明,miRNA326-5p 可与 DKC1 mRNA 结合抑制其蛋白表达,从而抑制 NB 细胞增殖,促进其凋亡,并调节凋亡蛋白的表达。
miRNA326-5p 靶向 DKC1 mRNA 调节凋亡相关蛋白,抑制神经母细胞瘤增殖,促进凋亡过程。