METHODS: HT29 and SW480 cells were cultured in media supplemented with L-OHP or 5-FU to establish drug-resistant strains. HT29 and SW480 drug-resistant cells were subcutaneously injected into the ventral nerves of nude mice at a dose of 5 × 106 to establish CRC drug-resistant animal models. CCK-8, Western blot, flow cytometry, Transwell and kit detection were used to detect the regulatory mechanism of energy metabolism reprogramming in drug-resistant CRC cells.
RESULTS: Compared with nonresistant strains, L-OHP/5-FU-resistant strains exhibited greater metabolic reprogramming. Functionally, SHP099 can restrain the metabolic reprogramming of L-OHP/5-FU-resistant strains and subsequently restrain the proliferation, colony formation, migration and spheroid formation of L-OHP/5-FU-resistant strains. Downstream mechanistic studies have shown that SHP099 interferes with the metabolic reprogramming of L-OHP/5-FU drug-resistant strains by suppressing the PI3K/AKT pathway, thereby restraining the malignant biological behavior of L-OHP/5-FU drug-resistant strains and alleviating CRC.
CONCLUSIONS: The combination of SHP099 can restrain the malignant biological behavior of L-OHP/5-FU-resistant CRC cells and alleviate the progression of CRC by interfering with the reprogramming of energy metabolism. This study explored the effect of SHP099 combination on dual-resistant CRC cells for the first time, and provided a new therapeutic idea for solving the problem of SHP099 insensitivity to CRC cells.
方法:将HT29和SW480细胞在补充L-OHP或5-FU的培养基中培养以建立耐药菌株。将HT29和SW480耐药细胞以5×106的剂量皮下注射到裸鼠腹侧神经中,建立CRC耐药动物模型。CCK-8,Western印迹,流式细胞术,采用Transwell和kit检测方法检测耐药CRC细胞能量代谢重编程的调控机制。
结果:与非耐药菌株相比,L-OHP/5-FU抗性菌株表现出更大的代谢重编程。功能上,SHP099可以抑制L-OHP/5-FU耐药菌株的代谢重编程,从而抑制其增殖,菌落形成,L-OHP/5-FU抗性菌株的迁移和球状体形成。下游机制研究表明,SHP099通过抑制PI3K/AKT通路干扰L-OHP/5-FU耐药菌株的代谢重编程,从而抑制L-OHP/5-FU耐药菌株的恶性生物学行为,缓解CRC。
结论:SHP099联合应用可通过干扰能量代谢的重编程,抑制L-OHP/5-FU耐药CRC细胞的恶性生物学行为,减轻CRC的进展。本研究首次探索SHP099联合对双重耐药CRC细胞的作用,为解决SHP099对CRC细胞不敏感的问题提供了新的治疗思路。