Abstract：
OBJECTIVE: Colorectal cancer (CRC) is one of the most common malignancies in China. At present, there is a problem that the CRC treatment drugs SHP099, L-OHP and 5-FU are insensitive to tumor cells. Combination medication is an important means to solve the insensitivity of medication alone. The purpose of this project was to explore the effect and molecular mechanism of SHP099 combination on the malignant biological behavior of L-OHP/5-FU resistant strains of CRC.
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.
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.
摘要：
目的：大肠癌（CRC）是中国最常见的恶性肿瘤之一。目前,存在CRC治疗药物SHP099、L-OHP和5-FU对肿瘤细胞不敏感的问题。联合用药是解决单独用药不敏感的重要手段。本课题旨在探讨SHP099组合对CRC耐药株L-OHP/5-FU恶性生物学行为的影响及分子机制。
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。
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。
