DataSheet1_Role of anoikis-related gene PLK1 in kidney renal papillary cell carcinoma: a bioinformatics analysis and preliminary verification on promoting proliferation and migration.docx

Background: Kidney renal papillary cell carcinoma (KIRP) is a rare malignancy with a very poor prognosis. Anoikis is a specific form of apoptosis involved in carcinogenesis, but the role of anoikis in KIRP has not been explored.
Methods: Anoikis-related genes (ARGs) were obtained from the GeneCards database and Harmonizome database and were used to identify different subtypes of KIRP and construct a prognostic model of KIRP. In addition, we also explored the immune microenvironment and enrichment pathways among different subtypes by consensus clustering into different subtypes. Drug sensitivity analysis was used to screen for potential drugs. Finally, we verified the mRNA and protein expression of the independent prognostic gene PLK1 in patient tissues and various cells and further verified the changes in relevant prognostic functions after constructing a PLK1 stable knockdown model using ShRNA.
Results: We identified 99 differentially expressed anoikis-related genes (DEGs) associated with KIRP survival, and selected 3 genes from them to construct a prognostic model, which can well predict the prognosis of KIRP patients. Consensus clustering divided KIRP into two subtypes, and there was a significant difference in survival rates between the two subtypes. Immune profiling revealed differing immune statuses between the two subtypes, and functional analysis reveals the differential activity of different functions in different subtypes. Drug sensitivity analysis screened out 15 highly sensitive drugs in the high-risk group and 11 highly sensitive drugs in the low-risk group. Univariate and multivariate Cox regression analysis confirmed that PLK1 was an independent prognostic factor in KIRP, and its mRNA and protein expression levels were consistent with gene differential expression levels, both of which were highly expressed in KIRP. Functional verification of PLK1 in KIRP revealed significant results. Specifically, silencing PLK1 inhibited cell proliferation, clonogenicity, and migration, which indicated that PLK1 plays an important role in the proliferation and migration of KIRP.
Conclusion: The prognosis model constructed by ARGs in this study can accurately predict the prognosis of KIRP patients. ARGs, especially PLK1, play an important role in the development of KIRP. This research can help doctors provide individualized treatment plans for KIRP patients and provide researchers with new research ideas.

背景：肾乳头状肾细胞癌（Kidney renal papillary cell carcinoma, KIRP）是一种预后极差的罕见恶性肿瘤。失巢凋亡（Anoikis）是一类参与肿瘤发生发展的特异性细胞凋亡形式，但目前尚未有研究阐明其在KIRP中的作用。
方法：本研究从GeneCards数据库与Harmonizome数据库获取失巢凋亡相关基因（Anoikis-related genes, ARGs），以此识别KIRP的不同分子亚型并构建KIRP预后预测模型。此外，通过一致性聚类将样本划分为不同亚型后，本研究还分析了各亚型间的免疫微环境与富集通路差异。采用药物敏感性分析筛选潜在治疗候选药物。最后，本研究在患者组织及多种细胞系中验证了独立预后因子Polo样激酶1（PLK1）的mRNA与蛋白表达水平，并通过构建短发夹RNA（ShRNA）介导的PLK1稳定敲低细胞模型，进一步验证了相关预后功能的变化。
结果：本研究共筛选出99个与KIRP生存相关的失巢凋亡相关差异表达基因（differentially expressed genes, DEGs），并从中选取3个基因构建预后预测模型，该模型可精准预测KIRP患者的预后情况。一致性聚类将KIRP样本划分为两个分子亚型，且两亚型间患者总体生存率存在显著差异。免疫特征分析显示两亚型的免疫微环境状态存在明显异质性，功能富集分析则揭示了不同亚型间功能活性的差异。药物敏感性分析分别在高风险组与低风险组中筛选出15种与11种高敏感性药物。单因素与多因素Cox回归分析证实，PLK1是KIRP的独立预后因子，其mRNA与蛋白表达水平与基因差异表达结果一致，均在KIRP组织中呈高表达状态。针对KIRP中PLK1的功能验证实验获得了显著性结果：沉默PLK1可显著抑制肿瘤细胞增殖、克隆形成能力与迁移能力，表明PLK1在KIRP的增殖与迁移过程中发挥关键调控作用。
结论：本研究构建的失巢凋亡相关基因预后预测模型可精准预测KIRP患者的预后情况。失巢凋亡相关基因，尤其是PLK1，在KIRP的发生发展进程中发挥重要调控作用。本研究可为临床医生为KIRP患者制定个体化治疗方案提供理论参考，同时也为相关领域研究者提供了全新的研究思路。