Objective  To explore the influence and mechanism of mechanistic target of rapamycin kinase (mTOR)/ receptor of advanced glycation end products (RAGE) pathway mediated-ferritinophagy on high glucose consumption promoting invasion and migration of colorectal cancer (CRC). Methods ① Patients and tissue samples. Clinical data and tissues were collected from CRC patients underwent surgery and completed the dietary questionnaire in the Second Affiliated Hospital of Harbin Medical University from October 2022 to October 2023. Real-time quantitative reverse transcription PCR (qRT-PCR) was used to analyzed the expression of nuclear receptor coactivator 4 (NCOA4) and ferritin in CRC and para-carcinoma tissues. ② Cell culture and treatment. The HT29 and HCT116 cells were treated by RPMI1640 medium containing 0, 35, 70, 105, 140 mmol/L glucose, and cell counting kit-8 (CCK-8) and lactate dehydrogenase (LDH) activity analysis were performed to confirm 105 mmol/L glucose was the optimal concentration in the current study. Then the HT29 and HCT116 cells were randomly divided into: control group, glucose group; control group, glucose group, si-RAGE group, and glucose+si-RAGE group; control group, glucose group, rapamycin group, and glucose+rapamycin group. Untreated HT29 and HCT116 cells were considered as control group. The cells in glucose group were treated with 105 mmol/L glucose for 48 h. The CRC cells in the si-RAGE group were transfected with si-RAGE for 6 h. The CRC cells in the rapamycin group were treated with 10 nmol/L rapamycin for 48 h. The CRC cells in the glucose+si-RAGE group were treated with 105 mmol/L glucose for 48 h combination transfected with si-RAGE for 6 h. The CRC cells in the glucose+rapamycin group were treated with 105 mmol/L glucose for 48 h combination treated with 10 nmol/L rapamycin for 48 h. Then electron microscopy and Western blot, wound healing assay and transwell assay were exhibited, respectively. ③ Azoxymethane (AOM)-induced CRC rat model. The effects of glucose consumption on malignant behavior and ferritinophagy mediated by mTOR/RAGE pathway were evaluated in AOM-induced CRC rat models. A total of 16 rats were randomly divided into control group and glucose group, the CRC number was recorded and HE staining of CRC tissues was further performed. The expression of RAGE, mTOR, NCOA4, and ferritin in colorectal tissues of rats from each group was detected by RT-qPCR. Results ① More lymphatic node metastasis and TNM Ⅲ/Ⅳ stages were observed in CRC patients from high glucose consumption group (P=0.004, P=0.004). Moreover, we confirmed that NCOA4 expression was significantly decreased (P<0.001) while ferritin was significantly increased (P<0.001) in CRC tissues especially in the CRC tissues from patients with positive lymph nodes metastasis. ② High glucose treatment significantly decreased autophagosomes in HT29 and HCT116 cells while si-RAGE transfection increased autophagic vacuoles compared to the control group. When compared with the glucose group, autophagosomes were increased in the glucose+si-RAGE group. Moreover, compared to the control group, the expressions of RAGE, p-mTOR, and ferritin were increased (P<0.001) while the expression of NCOA4 was decreased (P<0.001) in glucose group, but the expressions of RAGE, p-mTOR, and ferritin were decreased (P<0.001) while the expression of NCOA4 was increased (P<0.001) in the si-RAGE group; when compared with the glucose group, the expressions of RAGE, p-mTOR, and ferritin were downregulated (P<0.001) while the expression of NCOA4 was upregulated (P<0.001) in HT29 and HCT116 cells from the glucose+si-RAGE group. Compared to the control group, the HT29 and HCT116 cells in the glucose group performed enhanced wound scratch healing , migration and invasion viabilities (P<0.05); but the HT29 and HCT116 cells in the si-RAGE group performed impaired wound scratch healing, migration and invasion viabilities (P<0.05). When compared with the glucose group, the HT29 and HCT116 cells in the glucose+si-RAGE group performed impaired wound scratch healing, migration and invasion viabilities (P<0.05). ③ Rapamycin treatment significantly inhibited the expression of ferritin (P<0.05) but induced the expression of NCOA4 (P<0.05) compared to the control group. When compared with the glucose group, the expression of ferritin was downregulated (P<0.05) while the expression of NCOA4 was upregulated (P<0.05) in HT29 and HCT116 cells from the glucose+rapamycin group. Additionally, compared to the control group, rapamycin treatment performed inhibited effect on wound scratch healing, migration and invasion viabilities in the HT29 and HCT116 cells (P<0.05); while the HT29 and HCT116 cells in the glucose+rapamycin group performed impaired wound scratch healing, migration and invasion viabilities (P<0.05) when compared with the glucose group. ④ In the AOM induced CRC rat models, we found the more CRCs, aggravated cellular pleomorphism and upregulated expressions of RAGE, mTOR, ferritin (P<0.05) while downregulated expression of NCOA4 (P<0.05) in the glucose group than those of the control group. Conclusion High glucose consumption promote invasion and migration in CRC through suppressing ferritinophagy via activating the mTOR/RAGE pathway.

Citation： XIONG Xiao, CHEN Hao, LI Yuanliang, TANG Yan, ZHANG Haogang, QIAO Pengfei. High glucose consumption promoted invasion and migration in colorectal cancer through suppressing ferritinophagy via activating the RAGE/mTOR pathway. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2025, 32(4): 493-504. doi: 10.7507/1007-9424.202405003 Copy

Copyright ? the editorial department of CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY of West China Medical Publisher. All rights reserved