LINC00355 通过调控 Wnt/β-catenin 信号通路抑制胃癌细胞凋亡并促进其增殖。
LINC00355 inhibits apoptosis and promotes proliferation of gastric cancer cells by regulating Wnt/β-catenin signaling pathway.
机构信息
Department of General Surgery, Yantaishan Hospital, Yantai, China.
出版信息
Eur Rev Med Pharmacol Sci. 2020 Aug;24(16):8377-8383. doi: 10.26355/eurrev_202008_22634.
OBJECTIVE
To study the expression and biological functions of long intergenic non-protein coding ribonucleic acid 00355 (LINC00355) in gastric cancer (GC), and to explore its potential mechanism of action.
PATIENTS AND METHODS
The relative expression level of LINC00355 in 48 cases of GC tissues, the corresponding paracancerous tissues, and GC cells was determined using quantitative reverse transcription-polymerase chain reaction (qRT-PCR), and the interference efficiency of small interfering (si)-LINC00355 was detected via qRT-PCR. After knock-down of LINC00355, methyl thiazolyl tetrazolium (MTT) and 5-ethynyl-2'-deoxyuridine (EdU) assays were performed to detect the changes in the proliferation ability of GC cells, and the changes in the GC cell cycle distribution and apoptosis rate were examined by flow cytometry. Besides, Western blotting was conducted to verify the changes in the downstream signaling pathway of LINC00355.
RESULTS
Among 48 cases of GC tissues, there were 42 (87.5%) cases of LINC00355 expression up-regulation, and 6 (12.5%) cases of LINC00355 expression down-regulation. The qRT-PCR results revealed that the expression of LINC00355 was raised in 4 kinds of GC cells. After interference with LINC00355 expression, the MTT assay results indicated that the cell proliferation ability was inhibited, consistent with the EdU assay results. After LINC00355 was knocked down in GC cells, GC cells in experiment group had a higher apoptosis rate than those in si-NC group and arrested in the gap 0 (G0)/G1 phase. Moreover, it was found through Western blotting that the expressions of the molecular markers in the downstream wingless-INT (Wnt)/β-catenin signaling pathway were downregulated after interference with the expression of LINC00355.
CONCLUSIONS
LINC0035 exhibits an up-regulated expression in GC and regulates the Wnt/β-catenin signaling pathway to promote proliferation and inhibit apoptosis.
目的
研究长链非编码 RNA 00355（LINC00355）在胃癌（GC）中的表达和生物学功能，并探讨其潜在的作用机制。
方法
采用实时荧光定量聚合酶链反应（qRT-PCR）检测 48 例 GC 组织、相应癌旁组织和 GC 细胞中 LINC00355 的相对表达水平，并通过 qRT-PCR 检测小干扰（si）-LINC00355 的干扰效率。敲低 LINC00355 后，通过噻唑蓝（MTT）和 5-乙炔基-2'-脱氧尿苷（EdU）检测 GC 细胞增殖能力的变化，通过流式细胞术检测 GC 细胞周期分布和凋亡率的变化。此外，通过 Western blot 验证 LINC00355 下游信号通路的变化。
结果
在 48 例 GC 组织中，有 42 例（87.5%）LINC00355 表达上调，6 例（12.5%）LINC00355 表达下调。qRT-PCR 结果显示，4 种 GC 细胞中 LINC00355 的表达均升高。干扰 LINC00355 表达后，MTT 检测结果表明细胞增殖能力受到抑制，与 EdU 检测结果一致。在 GC 细胞中敲低 LINC00355 后，实验组 GC 细胞的凋亡率高于 si-NC 组，且停滞在 G0/G1 期。此外，通过 Western blot 发现，干扰 LINC00355 表达后，下游 Wnt/β-catenin 信号通路的分子标志物表达下调。
结论
LINC0035 在 GC 中呈上调表达，并通过调节 Wnt/β-catenin 信号通路促进增殖，抑制凋亡。
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