ENST00000428100通过MACC1调节mTOR通路影响结肠癌干细胞代谢可塑性的机制研究

Dysfuntional cellular metabolism exists in tumor cells. Cancer stem cells (CSC) have ability of self-renewal, which plays a critical role in tumor recurrence and metastasis. However, the metabolic pattern of colorectal CSC is largely unknown. Whether it is mainly based on the Warburg effect, or mitochondrial oxidative phosphorylation (OXPHOS) needs to be elucidated. Our previous study suggested that ENST00000428100, a long noncoding RNA (lncRNA), was involved in the sphere formation of colorectal CSC and glycolysis. Moreover, ENST00000428100 regulated the level of MACC1 and mTOR signaling pathway. Given that both MACC1 and mTOR pathway participated in glycolysis, this study will clarify the mechanism of ENST00000428100 regulating mTOR signaling pathway by MACC1 and determine the metabolic plasticity of colorectal CSC. First, we decide to demonstrate the metabolic pattern of colorectal CSC. Second, we want to detect the effects of ENST00000428100 and MACC1 on the metabolic pattern and plasticity of colorectal CSC. Third, we decide to understand the regulation between ENST00000428100 and MACC1, as well as their modulation of mTOR signaling pathway. Finally, we want to demonstrate the expression levels of ENST00000428100 and MACC1 in colorectal cancers, as well as their clinical significance. The purpose of this study was to clarify the regulation of metabolic plasticity in colorectal CSC, and the underlying mechanism of ENST00000428100-MACC1-mTOR network. Our study will provide new potential strategies for cancer diagnosis and treatment.

肿瘤细胞存在代谢异常。肿瘤干细胞具有自我更新能力，是肿瘤复发转移的关键因素。那么结肠癌干细胞的代谢模式是以Warburg效应为主，还是以线粒体氧化磷酸化为主，目前尚不清楚。前期发现，非编码RNA ENST00000428100（lncRNA）参与结肠癌干细胞形成和糖酵解，并调控MACC1和mTOR通路。基于MACC1和mTOR通路均参与肿瘤细胞糖酵解，本项目拟探讨lncRNA通过MACC1调节mTOR通路影响结肠癌干细胞代谢可塑性的机制。首先，明确结肠癌干细胞的能量代谢模式；其次，阐明lncRNA与MACC1对结肠癌干细胞能量代谢方式及其代谢可塑性的调节；再次，明确两分子间的调控方式，和对mTOR通路的作用；最后，明确上述分子在肿瘤组织中表达的临床意义。本研究旨在阐明以lncRNA-MACC1-mTOR为核心的结肠癌肿瘤干细胞代谢可塑性调控方式，并为肿瘤诊疗提供新的策略，及可能的干预措施。

本项目进展过程中围绕着“ENST00000428100通过MACC1调节mTOR通路影响结肠癌干细胞代谢可塑性的机制研究”开展了一系列研究工作。工作进行期间，首先，明确ENST00000428100及MACC1均参与mTOR通路调节，以及肿瘤细胞的糖代谢过程。同时，ENST00000428100低表达可抑制结直肠癌细胞增殖，促进细胞凋亡，减弱细胞转移潜能。在研究过程中明确了miR-200c介导KIF14通过AKT信号通路促进结直肠癌增殖的分子机制。同时，明确了非编码RNA circ0082007（circMET）通过与c-met竞争相关miR410-3p共同促进结直肠细胞增殖的作用。此外，通过对结直肠癌组织肿瘤干细胞进行单细胞测序，发现同一个病人病灶的肿瘤细胞在DNA水平存在异质性。.综上，本项目明确了非编码RNA ENST00000428100、miR-200c以及circ0082007对结直肠癌发生发展的影响以及作用方式。同时对肿瘤组织分离干细胞单细胞测序的研究，明确了肿瘤干细胞在肿瘤组织的异质性。该研究为深入了解非编码RNA及肿瘤干细胞在肿瘤发生发展中的作用及相关分子机制奠定了一定基础，具有一定的潜在应用价值。