Methionine restriction enhances the chemotherapeutic sensitivity of colorectal cancer stem cells by miR-320d/c-Myc axis
Methionine restriction enhances the chemotherapeutic sensitivity of colorectal cancer stem cells by miR-320d/c-Myc axis
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DOI：
10.1007/s11010-022-04416-1
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发表时间：
2022-04-08
影响因子：
4.3
通讯作者：
Xin，Lin
中科院分区：
文献类型：
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作者：
Liu，Chuan;Wang，Jin-Liang;Xin，Lin
结直肠癌干细胞（CRC-SCs）的化疗耐药已成为癌症临床治疗的重大挑战。蛋氨酸限制（MR）可增强化疗药物的治疗效果。本研究的目的是探讨MR影响CRC-SCs化疗敏感性的分子途径。通过磁激活细胞分选 (MACS) 分离 CD133+ 和 CD133–SW480 或 SW620 细胞。通过皮下接种CD133+SW480建立小鼠异种移植瘤模型。 MTT法用于检测细胞活力。流式细胞术检测细胞周期的时相分布。 Western blotting检测耐药相关蛋白表达。通过qRT-PCR检测miR-320d和转录因子c-Myc的表达。通过荧光素酶测定验证了 miR-320d 和 c-Myc 之间的相互作用。 CD133+SW480 和 SW620 细胞比 CD133-细胞对 5-氟尿嘧啶 (5-FU) 具有更强的耐药性。体外和体内实验表明，与单独给药相比，5-FU和MR联合治疗进一步抑制CD133+细胞活性和ATP结合盒亚家族G成员2（ABCG2）表达，并缩小肿瘤体积。干扰 miR-320d 或过度表达 c-Myc 可逆转 5-FU 和 MR 协同治疗引起的 CRC-SC 化疗敏感性增加。 miR-320d 可以靶向并调节 c-Myc。干扰 c-Myc 可以逆转由 miR-320d 下调引起的细胞活力和 ABCG2 表达的增加。总之, MR联合化疗可通过上调miR-320d抑制c-Myc表达来增强CRC-SCs的化疗敏感性, 为MR调控CRC-SCs的化疗敏感性奠定了分子基础。
Chemotherapy resistance of colorectal cancer stem cells (CRC-SCs) has become a major challenge in clinical treatment of cancer. Methionine restriction (MR) enhances the therapeutic effect of chemotherapeutic agents. The aim of this study was to explore the molecular pathways that MR affects the chemotherapeutic sensitivity of CRC-SCs. CD133+and CD133–SW480 or SW620 cells were isolated by magnetic-activated cell sorting (MACS). Mouse xenograft tumor model was established by subcutaneous inoculation of CD133+SW480. MTT assay was used to detect cell viability. Phase distribution of cell cycle was detected by flow cytometry. Western blotting was used to detect drug-resistant related protein expression. miR-320d and transcription factor c-Myc expressions were detected by qRT-PCR. The interaction between miR-320d and c-Myc was verified by luciferase assay. CD133+SW480 and SW620 cells were more resistant to 5-fluorouracil (5-FU) than CD133–cells. In vitro and in vivo experiments showed that 5-FU and MR combined therapy further inhibited CD133+cell activity and ATP binding cassette subfamily G member 2 (ABCG2) expression, and reduced tumor volume compared with drug administration alone. Interference with miR-320d or overexpression of c-Myc reversed the increased chemotherapeutic sensitivity of CRC-SCs induced by synergistic therapy with 5-FU and MR. miR-320d can target and regulate c-Myc. Interference with c-Myc could reverse the increase in cell viability and ABCG2 expression caused by down-regulation of miR-320d. In conclusion, the combined chemotherapy with MR can enhance the chemotherapeutic sensitivity of CRC-SCs by up-regulation of miR-320d to inhibit c-Myc expression, which lays a molecular basis for MR regulation of chemotherapeutic sensitivity of CRC-SCs.