（宫颈）癌前病变的Warburg-like effect与糖代谢重编程机制研究

Glycolysis is an important metabolic process of glucose. One of main metabolic characteristics of tumor cells is the high level of glycolysis even in the presence of oxygen, known as aerobic glycolysis or the Warburg effect. It remains unclear whether the Warburg effect causes tumor or the tumor exists the Warburg effect. Thus role of the Warburg effect in the cell malignant transformation or tumor development is key scientific question. Study of glucose metabolic reprogramming or the Warburg-like effect in premalignant tissues is helpful to answer this question. In previous study, we found that the key enzymes of glycolysis and glucose transporters are upregulated in high-grade squamous intraepithelial lesion (HSIL), suggesting that glucose metabolic reprogramming or the Warburg-like effect may occur in the early stage of the cell malignant transformation. In the current project, we will use the HSIL, a typical premalignant lesion, as a model to analyze characteristics of glucose metabolome and energy metabolism, detect the expression of key enzymes of glycolysis and glucose transporters, and investigate the regulation of the Warburg effect and the effect of HPV encoded oncoproteins on the Warburg effect. The data will uncover a Warburg-like effect in premalignant stage and provide a basis for the Warburg effect-based prevention and therapy of tumors.

糖酵解是葡萄糖代谢的一个重要过程。肿瘤细胞糖酵解活性增强，表现为有氧酵解即Warburg effect（瓦伯格效应），是肿瘤细胞代谢的一个显著特征，但不清楚这一现象是肿瘤发生的原因或结果，其在细胞恶性转化和肿瘤发生中的意义是肿瘤代谢研究的一个关键科学问题。调查癌前病变阶段是否存在糖代谢的重编程或瓦伯格样效应有助于回答这一问题。我们前期研究发现，宫颈癌前病变组织糖酵解关键酶及葡萄糖转运蛋白水平显著升高，提示细胞恶性转化早期已经开始糖代谢重编程，可能出现了类似瓦伯格效应样的代谢变化。本项目将以宫颈高度鳞状上皮内病变这一典型的癌前病变为模型，系统分析其葡萄糖代谢组改变及能量代谢特点，检测葡萄糖转运蛋白及糖酵解关键酶的表达变化，研究瓦伯格效应关键信号通路的调控并探讨HPV癌基因产物的影响，揭示在癌前病变阶段存在瓦伯格样效应，为通过干预瓦伯格效应进行肿瘤防治提供新的理论依据。

肿瘤细胞经历了代谢重编程。大多数肿瘤都存在糖酵解增强，即使在有氧气的情况下，这种现象被称为有氧糖酵解或Warburg effect。代谢重编程促进癌症的发展，甚至与肿瘤的发生有关。然而，尚不清楚观察到的代谢变化是癌症发展的驱动因素还是伴发现象。本项目应用代谢组学方法分析宫颈癌前病变和口腔癌前细胞的代谢特点，并用RP-PCR和western blot检测糖酵解酶在宫颈癌前病变中的表达。我们在口腔细胞和宫颈组织中分别鉴定出115种和23种具有可靠信号的代谢物。我们研究发现，组织细胞的代谢组可以明确区分宫颈癌前病变与正常宫颈上皮和宫颈癌。与正常宫颈上皮相比，宫颈癌前病变组织的葡萄糖消耗和乳酸分泌增加。与正常宫颈上皮比较，在宫颈癌前病变组织中糖酵解酶LDHA、HKⅡ和PKM2等的表达有增高趋势。根据细胞的代谢组特征，口腔癌前细胞DOK可与正常人口腔上皮细胞（HOEC）及口腔鳞癌细胞相区分。在DOK和HOEC之间鉴定出丙酮酸、谷氨酰胺、蛋氨酸、赖氨酸等4种关键的差异代谢产物。我们的研究结果表明，细胞代谢可能在肿瘤发生的早期阶段发生了向Warburg-like effect的重编程，提示代谢重编程在肿瘤发生发展中可能发挥重要作用。