Lamichhane Santosh, Mo Ji-Su, Sharma Grinsun, Joung Sun-Myoung, Chae Soo-Cheon
Department of Pathology, School of Medicine, Wonkwang University Iksan, Chonbuk 54538, Korea.
Digestive Disease Research Institute, Wonkwang University Iksan, Chonbuk 54538, Korea.
Am J Cancer Res. 2022 Jul 15;12(7):3223-3241. eCollection 2022.
The human microRNA 133A () was identified as a CRC-associated miRNA. It was down-regulated in human CRC tissues. We identified the putative and target genes by comparing the transcriptome analysis data of and knock-in cells with the candidate target genes predicted by bioinformatics tools. We identified 29 and 33 putative and direct target genes, respectively. Among them, we focused on the master transcription regulator gene SRY-box transcription factor 9 (), which exhibits a pleiotropic role in cancer. We confirmed that is a direct target gene of by luciferase reporter assay, quantitative RT-PCR, and western blot analysis. Overexpression of in CRC cell lines significantly decreased SOX9 and its downstream PIK3CA-AKT1-GSK3B-CTNNB1 and KRAS-BRAF-MAP2K1-MAPK1/3 pathways and increased apoptosis. Furthermore, functional studies reveal that cell proliferation, colony formation, and migration ability were significantly decreased by -overexpressed CRC cell lines. Knockdown of SOX9 in CRC cell lines by gene silencing showed similar results. We also used a xenograft model to show that overexpression suppresses tumor growth and proliferation. Our results suggest that regulates cell proliferation, migration, and apoptosis by targeting in human colorectal cancer.
人类微小RNA 133A（）被鉴定为与结直肠癌相关的微小RNA。它在人类结直肠癌组织中表达下调。通过将和敲入细胞的转录组分析数据与生物信息学工具预测的候选靶基因进行比较，我们鉴定出了假定的和靶基因。我们分别鉴定出了29个和33个假定的和直接靶基因。其中，我们重点关注了主要转录调节因子基因SRY盒转录因子9（），它在癌症中发挥着多效性作用。我们通过荧光素酶报告基因检测、定量逆转录-聚合酶链反应和蛋白质免疫印迹分析证实是 的直接靶基因。在结直肠癌细胞系中过表达显著降低了SOX9及其下游的PIK3CA-AKT1-GSK3B-CTNNB1和KRAS-BRAF-MAP2K1-MAPK1/3信号通路，并增加了细胞凋亡。此外，功能研究表明，过表达的结直肠癌细胞系的细胞增殖、集落形成和迁移能力显著降低。通过基因沉默在结直肠癌细胞系中敲低SOX9也显示出类似的结果。我们还使用异种移植模型表明过表达抑制肿瘤生长和增殖。我们的结果表明，在人类结直肠癌中通过靶向调节细胞增殖、迁移和凋亡。
