Downregulation of VRK1 Inhibits Progression of Lung Squamous Cell Carcinoma through DNA Damage.

IF 2.1 4区 医学 Q3 RESPIRATORY SYSTEM Canadian respiratory journal Pub Date : 2023-01-01 DOI:10.1155/2023/4533504
Ning Du, Boxiang Zhang, Yunfeng Zhang
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Abstract

Background: Lung squamous cell carcinoma (LUSC) is a common malignancy. And the antitumor effect of bovine pox virus-associated kinase 1 (VRK1) is becoming a hot research topic.

Methods: VRK1 expression and prognosis in LUSC were analyzed using the GEPIA database. The expression of VRK1 mRNA was detected in 25 LUSC clinical tissue samples by RT-PCR. VRK1 shRNA was transfected into LUSC NCI-H520 and SK-MES-1 cell lines to interfere with VRK1 expression, and the efficiency of VRK1 shRNA interference was detected by the western blot. The effects of VRK1 downregulation on LUSC cell viability, migration, cell cycle, and apoptosis were analyzed by the CCK8 assay, scratch assay, transwell assay, and flow cytometry. The effect of VRK1 downregulation on DNA damage response (DDR) was examined by immunofluorescence staining and western blot assays and further validated by in vivo experiments.

Results: VRK1 was highly expressed in both LUSC tissues and cells. Survival analysis showed that the overall survival of LUSC patients with high VRK1 expression was significantly lower than that of LUSC patients with low VRK1 expression (P=0.0026). The expression level of the VRK1 gene was significantly higher in cancer tissues of LUSC patients than in paracancerous tissues. After transfection of VRK1 shRNA in both LUSC cells, cell activity decreased (P < 0.001), migration ability started to be inhibited (P < 0.001), the ratio of G0/G1 phase cells increased (P < 0.001), and apoptosis rate increased (P < 0.001). Immunofluorescence and western blot results showed that shVRK1 increased the level of γ-H2A.X (P < 0.001) and promoted apoptosis of tumor cells (P < 0.001). In addition, the results of animal experiments showed that shVRK1 had antitumor effects (P < 0.001) and a combined effect with DOX (P < 0.001).

Conclusion: The downregulation of VRK1 significantly affected the proliferation, apoptosis, migration, and cell cycle progression of LUSC cells via DDR, suggesting that VRK1 is a suitable target for potential LUSC therapy.

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VRK1下调通过DNA损伤抑制肺鳞状细胞癌的进展
背景:肺鳞状细胞癌是一种常见的恶性肿瘤。而牛痘病毒相关激酶1 (VRK1)的抗肿瘤作用正成为研究的热点。方法:采用GEPIA数据库分析VRK1在LUSC中的表达及预后。RT-PCR检测25例LUSC临床组织样本中VRK1 mRNA的表达。将VRK1 shRNA转染到LUSC NCI-H520和SK-MES-1细胞系中,干扰VRK1的表达,并通过western blot检测VRK1 shRNA的干扰效率。通过CCK8实验、scratch实验、transwell实验和流式细胞术分析VRK1下调对LUSC细胞活力、迁移、细胞周期和凋亡的影响。通过免疫荧光染色和western blot检测VRK1下调对DNA损伤反应(DDR)的影响,并通过体内实验进一步验证VRK1下调对DDR的影响。结果:VRK1在LUSC组织和细胞中均有高表达。生存分析显示,VRK1高表达的LUSC患者的总生存率显著低于VRK1低表达的LUSC患者(P=0.0026)。VRK1基因在LUSC患者癌组织中的表达水平明显高于癌旁组织。两种LUSC细胞转染VRK1 shRNA后,细胞活性下降(P < 0.001),迁移能力开始受到抑制(P < 0.001), G0/G1期细胞比例增加(P < 0.001),凋亡率增加(P < 0.001)。免疫荧光和western blot结果显示,shVRK1使γ-H2A水平升高。X (P < 0.001),促进肿瘤细胞凋亡(P < 0.001)。此外,动物实验结果显示,shVRK1具有抗肿瘤作用(P < 0.001),并与DOX联合作用(P < 0.001)。结论:VRK1下调可通过DDR显著影响LUSC细胞的增殖、凋亡、迁移和细胞周期进程,提示VRK1是潜在的LUSC治疗靶点。
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来源期刊
Canadian respiratory journal
Canadian respiratory journal 医学-呼吸系统
CiteScore
4.20
自引率
0.00%
发文量
61
审稿时长
6-12 weeks
期刊介绍: Canadian Respiratory Journal is a peer-reviewed, Open Access journal that aims to provide a multidisciplinary forum for research in all areas of respiratory medicine. The journal publishes original research articles, review articles, and clinical studies related to asthma, allergy, COPD, non-invasive ventilation, therapeutic intervention, lung cancer, airway and lung infections, as well as any other respiratory diseases.
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