Zhang Haoran, Shen Zhanlong, Zhou Yushi, Zhang Zhen, Wang Quan, Zhang Mengmeng, Jiang Kewei, Wang Shan, Ye Yingjiang, Wang Bo
Department of Gastroenterological Surgery, Peking University People's Hospital, Beijing, China.
Laboratory of Surgical Oncology, Peking University People's Hospital, Beijing, China.
Front Genet. 2020 Sep 30;11:577948. doi: 10.3389/fgene.2020.577948. eCollection 2020.
MicroRNAs (miRNAs), such as miR-654-3p, regulate gene expression at the post-transcriptional level affecting malignant tumor behavior. However, the expression levels, function, and mechanism of miR-654-3p in colorectal cancer (CRC) are unknown.
The expression levels of miR-654-3p and SRC in 103 CRC tissues and matched normal colorectal tissues were detected by a quantitative real-time polymerase chain reaction (qRT-PCR). miR-654-3p was overexpressed by RNA mimics and SRC knockdown by siRNA. Function-based experiments were carried out to detect the proliferation and migration abilities in CRC cell lines. Flow cytometry assay was performed to evaluate the effect of miR-654-3p on cell apoptosis and cycle distribution. Xenograft tumor models in nude mice were utilized to evaluate miR-654-3p functions . Dual-fluorescence reporter assay was used to verify the direct binding between miR-654-3p and SRC.
miR-654-3p was downregulated in CRC tissues as compared to matched normal colorectal tissues. The expression levels of miR-654-3p were closely associated with distant metastasis. In addition, elevated expression of miR-654-3p in CRC patients prolonged the overall survival. Upregulated miR-654-3p significantly suppressed the proliferation and migration capacity of CRC cells by enhancing apoptosis and promoting G0/G1 phase arrest. The direct binding between miR-654-3p and SRC was verified by the dual-luciferase reporter gene. Furthermore, the suppression of proliferation and migration capacity by elevated miR-654-3p level could be reversed by overexpressing SRC.
miR-654-3p acts as a tumor suppressor through regulating SRC. It might also serve as a diagnostic and prognostic indicator and a novel molecular target for CRC therapy.
微小RNA（miRNA），如miR-654-3p，在转录后水平调节基因表达，影响恶性肿瘤行为。然而，miR-654-3p在结直肠癌（CRC）中的表达水平、功能及机制尚不清楚。
采用定量实时聚合酶链反应（qRT-PCR）检测103例CRC组织及配对的正常结直肠组织中miR-654-3p和SRC的表达水平。通过RNA模拟物过表达miR-654-3p，利用小干扰RNA（siRNA）敲低SRC。进行基于功能的实验以检测CRC细胞系中的增殖和迁移能力。采用流式细胞术分析评估miR-654-3p对细胞凋亡和细胞周期分布的影响。利用裸鼠异种移植瘤模型评估miR-654-3p的功能。采用双荧光素酶报告基因检测法验证miR-654-3p与SRC之间的直接结合。
与配对的正常结直肠组织相比，CRC组织中miR-654-3p表达下调。miR-654-3p的表达水平与远处转移密切相关。此外，CRC患者中miR-654-3p表达升高可延长总生存期。上调miR-654-3p可通过增强凋亡和促进G0/G1期阻滞显著抑制CRC细胞的增殖和迁移能力。双荧光素酶报告基因验证了miR-654-3p与SRC之间的直接结合。此外，过表达SRC可逆转miR-654-3p水平升高对增殖和迁移能力的抑制作用。
miR-654-3p通过调节SRC发挥肿瘤抑制作用。它还可能作为CRC诊断和预后的指标以及CRC治疗的新分子靶点。
