Department of Integrated Traditional & Western Medicine, Ningbo Haishu Traditional Chinese Medicine Hospital, Ningbo, Zhejiang, 315010, China; Department of Gastroenterology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310060, China.
Department of Gastroenterology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, 310060, China.
Phytomedicine. 2024 Oct;133:155906. doi: 10.1016/j.phymed.2024.155906. Epub 2024 Jul 25.
Colorectal cancer (CRC) and its chemoresistance pose significant threats to human health. Gegen Qinlian Decoction (GQD) is frequently employed alongside chemotherapy drugs for the treatment of CRC and various intestinal disorders. Despite its widespread use, there is limited research investigating the mechanisms through which GQD reverses chemoresistance.
This study investigated the mechanism by which GQD reverses oxaliplatin (OXA) resistance in CRC.
A YTH N6-methyladenosine RNA binding protein 1 (YTHDF1)-knockdown OXA-resistant cell line was constructed by lentivirus to clarify YTHDF1-mediated chemoresistance through the regulation of glutaminase 1 (GLS1). The efficacy of GQD in reversing OXA resistance in CRC in vitro was evaluated by Cell Counting Kit-8, western blotting, quantitative real-time polymerase chain reaction, and glutaminase activity assays. In vivo validation was performed by constructing tumor xenografts in nude mice with OXA-resistant cells. In addition, mouse feces were collected and a 16S rDNA assay was performed to assess the regulation of intestinal flora by GQD.
Overexpression of YTHDF1 upregulated GLS1 expression and induced OXA-resistance in CRC. GQD induced apoptosis in LoVo/OXAR, increased OXA accumulation in LoVo/OXAR, inhibited expression of YTHDF1 and GLS1 when administered alone and in combination with OXA, and suppressed GLS1 activity to reverse drug resistance with good synergistic effects. GQD and OXA combination or GLS1 inhibitor alleviated OXA toxicity, reduced the volume of tumor xenografts in nude mice, inhibited YTHDF1 and GLS1 protein expression and GLS1 activity, adjusted the intestinal flora, and significantly reversed the increased Firmicutes/Bacteroidetes ratio.
GQD has shown superior efficacy in reversing OXA-resistance and increasing sensitivity. These findings indicate that the therapy combined with GQD has potential utility in the treatment of OXA-resistant CRC.
结直肠癌（CRC）及其化疗耐药性对人类健康构成重大威胁。葛根芩连汤（GQD）常与化疗药物联合用于治疗 CRC 和各种肠道疾病。尽管 GQD 被广泛应用，但对于其逆转耐药性的机制研究却相对较少。
本研究旨在探讨 GQD 逆转 CRC 奥沙利铂（OXA）耐药的机制。
通过慢病毒构建 YTH N6-甲基腺苷 RNA 结合蛋白 1（YTHDF1）敲低的 OXA 耐药细胞系，阐明 YTHDF1 通过调节谷氨酰胺酶 1（GLS1）介导的化疗耐药性。通过细胞计数试剂盒-8、Western blot、实时定量聚合酶链反应和谷氨酰胺酶活性测定评估 GQD 在体外逆转 CRC 对 OXA 耐药的疗效。通过构建 OXA 耐药细胞的裸鼠肿瘤异种移植进行体内验证。此外，收集小鼠粪便进行 16S rDNA 检测，以评估 GQD 对肠道菌群的调节作用。
YTHDF1 的过表达上调了 GLS1 的表达，诱导了 CRC 的 OXA 耐药性。GQD 诱导 LoVo/OXAR 细胞凋亡，增加了 LoVo/OXAR 中 OXA 的积累，单独和联合 OXA 给药时抑制了 YTHDF1 和 GLS1 的表达，并抑制了 GLS1 活性以产生良好的协同逆转耐药作用。GQD 和 OXA 联合或 GLS1 抑制剂减轻了 OXA 的毒性，减少了裸鼠肿瘤异种移植的体积，抑制了 YTHDF1 和 GLS1 蛋白表达和 GLS1 活性，调节了肠道菌群，并显著逆转了增加的厚壁菌门/拟杆菌门比值。
GQD 在逆转 OXA 耐药和增加敏感性方面显示出优异的疗效。这些发现表明，联合 GQD 的治疗方法在治疗 OXA 耐药 CRC 方面具有潜在的应用价值。
