Department of Biotechnology and Laboratory Science in Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan.
Division of Pediatrics, Taipei City Hospital, Taipei, Taiwan.
Hum Gene Ther. 2024 Oct;35(19-20):855-867. doi: 10.1089/hum.2023.207. Epub 2024 Jul 4.
Early diagnosis and intervention are pivotal in reducing colorectal cancer (CRC) incidence and enhancing patient outcomes. In this study, we focused on three genes, AQP8, GUCA2B, and SPIB, which exhibit high coexpression and play crucial roles in suppressing early-stage CRC. Our objective was to identify key miRNAs that can mitigate CRC tumorigenesis and modulate the coexpression network involving these genes. We conducted a comprehensive analysis using large-scale tissue mRNA data from The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus to validate the coexpression of AQP8, GUCA2B, and SPIB, and to assess their diagnostic and prognostic significance in CRC. The mRNA-miRNA interactions were examined using MiRNet and the Encyclopedia of RNA Interactomes. Furthermore, using various molecular techniques, we conducted miRNA inhibitor transfection experiments in HCT116 cells to evaluate their effects on cell growth, migration, and gene/protein expression. Our findings revealed that, compared with normal tissues, AQP8, GUCA2B, and SPIB exhibited high coexpression and were downregulated in CRC, particularly during tumorigenesis. OncoMirs, hsa-miR-182-5p, and hsa-miR-27a-3p, were predicted to regulate these genes. MiRNA inhibition experiments in HCT116 cells demonstrated the inhibitory effects of miR-27a-3p and miR-182-5p on GUCA2B mRNA and protein expression. These miRNAs promoted the proliferation of CRC cells, possibly through their involvement in the GUCA2B-GUCY2C axis, which is known to promote tumor growth. While the expressions of AQP8 and SPIB were barely detectable, their regulatory relationship with hsa-miR-182-5p remained inconclusive. Our study confirms that hsa-miR-27a-3p and hsa-miR-182-5p are oncomiRs in CRC. These miRNAs may contribute to GUCY2C dysregulation by downregulating GUCA2B, which encodes uroguanylin. Consequently, hsa-miR-182-5p and hsa-miR-27a-3p show promise as potential targets for early intervention and treatment in the early stages of CRC.
早期诊断和干预是降低结直肠癌(CRC)发病率和改善患者预后的关键。在这项研究中,我们专注于三个基因,AQP8、GUCA2B 和 SPIB,它们表现出高度的共表达,并且在抑制早期 CRC 中起着关键作用。我们的目标是确定可以减轻 CRC 肿瘤发生并调节涉及这些基因的共表达网络的关键 miRNA。我们使用来自癌症基因组图谱(TCGA)和基因表达综合数据库的大规模组织 mRNA 数据进行了全面分析,以验证 AQP8、GUCA2B 和 SPIB 的共表达,并评估它们在 CRC 中的诊断和预后意义。使用 MiRNet 和 RNA 相互作用百科全书检查了 mRNA-miRNA 相互作用。此外,我们使用各种分子技术在 HCT116 细胞中进行 miRNA 抑制剂转染实验,以评估它们对细胞生长、迁移和基因/蛋白表达的影响。我们的研究结果表明,与正常组织相比,AQP8、GUCA2B 和 SPIB 在 CRC 中表现出高度的共表达并且下调,特别是在肿瘤发生期间。OncoMirs、hsa-miR-182-5p 和 hsa-miR-27a-3p 被预测调节这些基因。在 HCT116 细胞中的 miRNA 抑制实验表明,miR-27a-3p 和 miR-182-5p 对 GUCA2B mRNA 和蛋白表达具有抑制作用。这些 miRNA 促进 CRC 细胞的增殖,可能通过它们参与已知促进肿瘤生长的 GUCA2B-GUCY2C 轴。虽然 AQP8 和 SPIB 的表达几乎检测不到,但它们与 hsa-miR-182-5p 的调节关系仍不清楚。我们的研究证实 hsa-miR-27a-3p 和 hsa-miR-182-5p 是 CRC 中的癌基因。这些 miRNA 可能通过下调编码尿鸟苷素的 GUCA2B 导致 GUCY2C 失调。因此,hsa-miR-182-5p 和 hsa-miR-27a-3p 有望成为 CRC 早期阶段早期干预和治疗的潜在靶点。