National Center for Orthopaedics, Beijing Research Institute of Traumatology and Orthopaedics, Beijing JiShuiTan Hospital, Beijing, China.
State Key Laboratory of Molecular Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
PeerJ. 2024 Mar 18;12:e16817. doi: 10.7717/peerj.16817. eCollection 2024.
Antibody-based platforms (, ADC) have emerged as one of the most encouraging tools for the cancer resistance caused by cancer stem cells (CSCs) enrichment. Our study might provide a promising therapeutic direction against drug resistance and serve as a potential precursor platform for screening ADC.
The cell migration, invasion, drug resistance, and self-renewal were assessed by the cell invasion and migration assay, wound healing assay, CCK-8 assay, colony formation assay, and sphere formation assay, respectively. The expression profiles of CSCs (ALDH and CD44) subpopulations were screened by flow cytometry. The western blot and cell immunofluorescence assay were used to evaluate pathway-related protein expression in both anti-ENO1 antibody, MET combined with DPP/CTX-treated CSCs.
In the present study, western blot and flow cytometry verified that anti-ENO1 antibody target the CD44 subpopulation by inhibiting the PI3K/AKT pathway, while metformin might target the ALDH subpopulation through activation of the AMPK pathway and thus reverse drug resistance to varying degrees. Subsequently, investigation indicated that anti-ENO1 antibody, metformin combined with cisplatin/cetuximab could simultaneously target ALDH and CD44 subpopulations. The combination also inhibited the CSCs proliferation, migration, invasion, and sphere formation; which may result in overcoming the drug resistance. Then, molecular mechanism exploration verified that the anti-ENO1 antibody, metformin combined with cisplatin/cetuximab inhibited the Wnt/β-catenin signaling.
The study preliminarily revealed anti-ENO1 antibody combined with metformin could overcome drug resistance against CSCs by inhibiting the Wnt//β-catenin pathway and might serve as a potential precursor platform for screening ADC. More importantly, it is reasonably believed that antibody-based drug combination therapy might function as an encouraging tool for oncotherapy.
抗体为基础的平台（ADC）已经成为针对癌症干细胞（CSC）富集引起的癌症耐药性的最有前途的工具之一。我们的研究可能为对抗耐药性提供有希望的治疗方向，并作为筛选 ADC 的潜在前体平台。
通过细胞侵袭和迁移测定、划痕愈合测定、CCK-8 测定、集落形成测定和球体形成测定分别评估细胞迁移、侵袭、耐药性和自我更新。通过流式细胞术筛选 CSCs（ALDH 和 CD44）亚群的表达谱。使用 Western blot 和细胞免疫荧光测定评估抗 ENO1 抗体、MET 联合 DPP/CTX 处理的 CSCs 中通路相关蛋白的表达。
在本研究中，Western blot 和流式细胞术验证了抗 ENO1 抗体通过抑制 PI3K/AKT 通路靶向 CD44 亚群，而二甲双胍可能通过激活 AMPK 通路靶向 ALDH 亚群，从而在不同程度上逆转耐药性。随后，研究表明抗 ENO1 抗体、二甲双胍联合顺铂/西妥昔单抗可以同时靶向 ALDH 和 CD44 亚群。该组合还抑制了 CSCs 的增殖、迁移、侵袭和球体形成，从而可能克服耐药性。然后，分子机制探索验证了抗 ENO1 抗体、二甲双胍联合顺铂/西妥昔单抗抑制了 Wnt/β-catenin 信号通路。
该研究初步揭示了抗 ENO1 抗体联合二甲双胍通过抑制 Wnt//β-catenin 通路可以克服 CSCs 的耐药性，并可能作为筛选 ADC 的潜在前体平台。更重要的是，合理地认为抗体为基础的药物联合治疗可能成为肿瘤治疗的有希望的工具。
