Department of Gastroenterology, Shengjing Hospital of China Medical University, PR China.
Department of Pulmonary and Critical Care Medicine, Shengjing Hospital of China Medical University, PR China.
Mol Immunol. 2020 Dec;128:287-297. doi: 10.1016/j.molimm.2020.10.007. Epub 2020 Nov 25.
JAK/STAT pathway has been well confirmed in the development of colorectal cancer (CRC), however, the exact mechanism is unclear. Therefore, we aimed to identify key genes involved in JAK/STAT pathway in CRC, as well as the potential mechanism. RT² profiler PCR arrays were performed to identify key genes of the JAK/STAT pathway. GO, KEGG pathway and PPI analyses were performed to screen the main functions of differentially expressed genes (DEGs). Moreover, the expression of DEGs was detected by GEPIA based on TCGA database and verified by qPCR and/or Western blot. Subsequently, the association between the two DEGs (CXCL9 and IL6ST) and clinicopathological features were determined by immunohistochemistry, and survival analysis was also conducted. Finally, the effects of IL6ST overexpression on STAT3 activation and HT29 cell functions were analyzed. A total of 14 DEGs were identified. Among the DEGs, GHR, NR3C1, IL6ST and A2M were confirmed to be statistically decreased, while CXCL9 was significantly increased in the CRC tissues. Furthermore, CXCL9 was significantly associated with differentiation, lymph node metastasis, distant metastasis and invasion, while IL6ST was related with tumor size, differentiation, stage and invasion. Patients with high expression of IL6ST presented significantly lower lifetime, however, CXCL9 showed the opposite results without significance. Additionally, we found that overexpression of IL6ST statistically elevated p-STAT3 level, cell viability, adhesion rate and migration, and decreased apoptosis, but had no effects on cell cycle. Our results suggest that IL6ST is a critical key gene involved in JAK/STAT signaling pathway in CRC.
JAK/STAT 通路在结直肠癌（CRC）的发生发展中已得到充分证实，但其确切机制尚不清楚。因此，我们旨在鉴定 CRC 中 JAK/STAT 通路相关的关键基因，并探讨其潜在的作用机制。通过 RT² Profiler PCR 阵列筛选 JAK/STAT 通路的关键基因。GO、KEGG 通路和 PPI 分析筛选差异表达基因（DEGs）的主要功能。基于 TCGA 数据库，通过 GEPIA 检测 DEGs 的表达情况，并通过 qPCR 和/或 Western blot 进行验证。随后，通过免疫组化检测两个 DEG（CXCL9 和 IL6ST）与临床病理特征的相关性，并进行生存分析。最后，分析 IL6ST 过表达对 STAT3 激活和 HT29 细胞功能的影响。共筛选到 14 个 DEGs。其中，GHR、NR3C1、IL6ST 和 A2M 被证实统计学下调，而 CXCL9 在 CRC 组织中显著上调。此外，CXCL9 与分化、淋巴结转移、远处转移和浸润显著相关，而 IL6ST 与肿瘤大小、分化、分期和浸润相关。IL6ST 高表达的患者总生存期明显降低，但 CXCL9 表达与生存期的相关性无统计学意义。此外，我们发现 IL6ST 的过表达可显著升高 p-STAT3 水平、细胞活力、黏附率和迁移率，降低细胞凋亡率，但对细胞周期无影响。综上，我们的研究结果表明，IL6ST 是 CRC 中 JAK/STAT 信号通路的关键基因。
