Colorectal Cancer Center, Zhongshan Hospital, Fudan University, Shanghai, China.
Department of General Surgery, Zhongshan Hospital, Fudan University, Shanghai, China.
J Transl Med. 2022 Mar 3;20(1):106. doi: 10.1186/s12967-022-03308-8.
Colorectal cancer (CRC) is one of the lethal cancers with a high mortality rate worldwide and understanding the mechanisms behind its progression is critical for improving patients' prognosis and developing therapeutics. MiR-500a-3p has been demonstrated to be involved in the progression of several human cancers but its role in CRC remains unclear. The aim of this study is to uncover the expression pattern and mechanisms of action of miR-500a-3p during the CRC progression.
The expression of miR-500a-3p and Cyclin-dependent kinases 6 (CDK6) in 134 CRC tissues were tested by quantitative PCR (qPCR) and immunohistochemistry staining (IHC), respectively. The effect of miR-500a-3p on cell proliferation was explored in vitro and in vivo. The glycolysis of CRC cells was determined by Mass Spectrometry and Seahorse XF 96 Extracellular Flux Analyzer. A dual-luciferase reporter assay was performed to validate the relationship between miR-500a-3p and CDK6.
miR-500a-3p was abnormally downregulated in CRC tissues and cell lines and was negatively associated with a worse prognosis. miR-500a-3p mimics impeded CRC cell proliferation in vitro and in vivo. miR-500a-3p inhibited glucose consumption, lactate and ATP production, and down-regulated the expression of hexokinase2 (HK2). In silico prediction combined with western blot and luciferase assay confirmed that CDK6 is a direct target of miR-500a-3p. Overexpression of CDK6 phenotypically rescued the inhibitory effect of miR-500a-3p on the proliferation and glycolysis of CRC cells.
Our study revealed a potential tumor-suppressive role of miR-500a-3p in CRC, specifically targeting CDK6 and inhibiting cancer cell proliferation and aerobic glycolysis, which may provide new insights into novel prognostic biomarkers and therapeutic targets for CRC.
结直肠癌（CRC）是一种具有高死亡率的致命癌症，了解其进展的机制对于改善患者预后和开发治疗方法至关重要。miR-500a-3p 已被证明参与了几种人类癌症的进展，但它在 CRC 中的作用尚不清楚。本研究旨在揭示 miR-500a-3p 在 CRC 进展过程中的表达模式和作用机制。
通过定量 PCR（qPCR）和免疫组织化学染色（IHC）分别检测 134 例 CRC 组织中 miR-500a-3p 和细胞周期蛋白依赖性激酶 6（CDK6）的表达。在体外和体内研究 miR-500a-3p 对细胞增殖的影响。通过质谱和 Seahorse XF 96 细胞外通量分析仪测定 CRC 细胞的糖酵解。进行双荧光素酶报告基因实验验证 miR-500a-3p 和 CDK6 之间的关系。
miR-500a-3p 在 CRC 组织和细胞系中异常下调，与预后不良呈负相关。miR-500a-3p 模拟物在体外和体内抑制 CRC 细胞增殖。miR-500a-3p 抑制葡萄糖消耗、乳酸和 ATP 产生，并下调己糖激酶 2（HK2）的表达。通过计算机预测结合 Western blot 和荧光素酶实验证实 CDK6 是 miR-500a-3p 的直接靶标。CDK6 的过表达表型上挽救了 miR-500a-3p 对 CRC 细胞增殖和糖酵解的抑制作用。
本研究揭示了 miR-500a-3p 在 CRC 中具有潜在的肿瘤抑制作用，其特异性靶向 CDK6 并抑制癌细胞增殖和有氧糖酵解，这可能为 CRC 的新型预后生物标志物和治疗靶点提供新的见解。
Zotero 插件
