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 (). 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 (), migration ability started to be inhibited (), the ratio of G0/G1 phase cells increased (), and apoptosis rate increased (). Immunofluorescence and western blot results showed that shVRK1 increased the level of γ-H2A.X () and promoted apoptosis of tumor cells (). In addition, the results of animal experiments showed that shVRK1 had antitumor effects () and a combined effect with DOX (). 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.

中文翻译：

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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（LUSC）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热点。方法。使用 GEPIA 数据库分析 LUSC 中 VRK1 的表达和预后。采用RT-PCR方法检测25份LUSC临床组织样本中VRK1 mRNA的表达情况。将VRK1 shRNA转染LUSC NCI-H520和SK-MES-1细胞系干扰VRK1表达，通过western blot检测VRK1 shRNA干扰效率。通过 CCK8 实验、划痕实验、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蛋白质印迹分析检测 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。结果。VRK1 在 LUSC 组织和细胞中高表达。生存分析显示，VRK1高表达的LUSC患者的总生存率显着低于VRK1低表达的LUSC患者（）。LUSC患者癌组织中VRK1基因的表达量显着高于癌旁组织。在两种 LUSC 细胞中转染 VRK1 shRNA 后，细胞活性下降（），迁移能力开始受到抑制（）， G0/G1期细胞比例增加（），细胞凋亡率增加（）。免疫荧光和蛋白质印迹结果表明shVRK1增加了γ -H2A.X的水平（）并促进肿瘤细胞凋亡（）。此外，动物实验结果表明shVRK1具有抗肿瘤作用（）以及与 DOX 的综合作用（）。 结论。VRK1的下调通过DDR显着影响LUSC细胞的增殖、凋亡、迁移和细胞周期进程，表明VRK1是潜在LUSC治疗的合适靶点。