Department of Biochemistry and Molecular Biology, Chongqing Medical University, Chongqing 400016, China.
Molecular Medicine and Cancer Research Center, Chongqing Medical University, Chongqing 400016, China.
Genes (Basel). 2021 Jan 19;12(1):120. doi: 10.3390/genes12010120.
We previously demonstrated that proline-rich protein 11 (PRR11) and spindle and kinetochore associated 2 (SKA2) constituted a head-to-head gene pair driven by a prototypical bidirectional promoter. This gene pair synergistically promoted the development of non-small cell lung cancer. However, the signaling pathways leading to the ectopic expression of this gene pair remains obscure. In the present study, we first analyzed the lung squamous cell carcinoma (LSCC) relevant RNA sequencing data from The Cancer Genome Atlas (TCGA) database using the correlation analysis of gene expression and gene set enrichment analysis (GSEA), which revealed that the PRR11-SKA2 correlated gene list highly resembled the Hedgehog (Hh) pathway activation-related gene set. Subsequently, GLI1/2 inhibitor GANT-61 or GLI1/2-siRNA inhibited the Hh pathway of LSCC cells, concomitantly decreasing the expression levels of PRR11 and SKA2. Furthermore, the mRNA expression profile of LSCC cells treated with GANT-61 was detected using RNA sequencing, displaying 397 differentially expressed genes (203 upregulated genes and 194 downregulated genes). Out of them, one gene set, including BIRC5, NCAPG, CCNB2, and BUB1, was involved in cell division and interacted with both PRR11 and SKA2. These genes were verified as the downregulated genes via RT-PCR and their high expression significantly correlated with the shorter overall survival of LSCC patients. Taken together, our results indicate that GLI1/2 mediates the expression of the PRR11-SKA2-centric gene set that serves as an unfavorable prognostic indicator for LSCC patients, potentializing new combinatorial diagnostic and therapeutic strategies in LSCC.
我们之前证明，富含脯氨酸蛋白 11（PRR11）和纺锤体和着丝粒相关蛋白 2（SKA2）构成了一个由典型双向启动子驱动的头对头基因对。这个基因对协同促进了非小细胞肺癌的发展。然而，导致这个基因对异位表达的信号通路仍然不清楚。在本研究中，我们首先使用基因表达相关性分析和基因集富集分析（GSEA）分析了来自癌症基因组图谱（TCGA）数据库的肺鳞状细胞癌（LSCC）相关 RNA 测序数据，结果表明 PRR11-SKA2 相关基因列表高度类似于 Hedgehog（Hh）通路激活相关基因集。随后，GLI1/2 抑制剂 GANT-61 或 GLI1/2-siRNA 抑制 LSCC 细胞的 Hh 通路，同时降低 PRR11 和 SKA2 的表达水平。此外，用 GANT-61 处理的 LSCC 细胞的 mRNA 表达谱通过 RNA 测序进行检测，显示 397 个差异表达基因（203 个上调基因和 194 个下调基因）。其中，一个包括 BIRC5、NCAPG、CCNB2 和 BUB1 的基因集参与细胞分裂，与 PRR11 和 SKA2 相互作用。通过 RT-PCR 验证了这些基因是下调基因，它们的高表达与 LSCC 患者的总生存期较短显著相关。总之，我们的结果表明，GLI1/2 介导 PRR11-SKA2 为中心的基因集的表达，作为 LSCC 患者不利的预后指标，为 LSCC 提供新的组合诊断和治疗策略。
