KRAS基因能诱导小鼠支气管肺泡交界处干细胞（BASCs）恶性转化并发展为肺腺癌，然其对BASCs细胞恶性转化过程中能量代谢的影响尚缺乏研究。本项目组前期发现KRAS突变人肺腺癌干细胞采用有氧糖酵解的能量代谢模式，结合近期国外研究发现突变KRAS可增强癌细胞膜葡萄糖转运蛋白1（GLUT1）表达来促进有氧糖酵解，由此提出KRAS突变可以通过加强BASCs细胞糖代谢，促进其恶性转化而成为具有自我更新及无限增殖能力的肺腺癌干细胞。本课题拟通过K-ras G12D 小鼠作为在体模型研究KRAS突变肺腺癌细胞：①糖酵解和氧化磷酸化模式之间的转变频率、强度和方式；②KRAS激活与能量代谢模式转变之间的关系及调控机制；③抑制KRAS表达后细胞生物能量学行为变化。旨在探讨KRAS介导的能量代谢模式转变与干细胞恶性转化的关系，探明KRAS突变型人LCSCs形成过程中能量代谢模式转变规律。

The metabolism of cancer stem cells and its regulatory mechanism is still unclear. We have previously indentified a population of lung cancer stem cells (LCSCs) from lung cancer cell line A549 which is KRAS mutant, and we found that aerobic glycolysis is a major determinant of the stemness of LCSCs/A549. It has also been reported that the mutant KRAS can promote aerobic glycolysis by upregulate glucose transporter 1 (GLUT1). we propose that Kras is implicated in the malignant transformation of BASCs by manipulating glycometabolism. K-ras G12D mice are used as model to study the transformation of BASCs into LCSCs: 1. the balance between glycolysis and oxidative phsophorylation;2.KRAS activation and metabolism model transformation;3.the metabolism pattern after KRAS inhibition. The findings can help to explain the role of KRAS in controlling metabolism during BASCs malignant transformation.

KRAS基因编码一个具有GTP酶活性的RAS蛋白，是EGFR信号通路的关键调控蛋白，位于EGFR受体下游，可促进细胞增殖、迁移、生存、血管新生、代谢和免疫逃逸。本项目组前期发现KRAS突变人肺腺癌干细胞采用有氧糖酵解的能量代谢模式，提出K-ras基因可以通过影响能量代谢途径调控肺癌细胞增殖可能。本项目在前期工作的基础上，采用特异性药物抑制剂及特异性 shRNA，体外模型明确了K-ras通过作用于HK2、PKM2影响肺癌细胞有氧糖酵解，K-ras与HK2、PKM2表达呈正相关；通过临床标本组织芯片相关性分析验证K-ras与HK2、PKM2的关系；采用HK2特异性抑制剂抑制HK2活性，研究对K-ras突变肺癌细胞增殖、凋亡的影响及其逆转顺铂耐药、EGFR-TKI耐药的作用。通过本项目，不但能明确k-ras基因对肺癌细胞能量代谢途径的影响，而且也将为临床肺癌治疗耐药提供新思路、新靶点。