Xu Chuchu, Zhu Renjun, Dai Qingfeng, Li Yaoqing, Hu Gengyuan, Tao Kelong, Xu Yuhong, Xu Guangen, Zhang Guolin
Department of Gastrointestinal Surgery, Shaoxing People's Hospital, Shaoxing, Zhejiang Province, China.
Department of Emergency, Shaoxing People's Hospital, Shaoxing, Zhejiang Province, China.
J Cell Mol Med. 2025 Mar;29(6):e70470. doi: 10.1111/jcmm.70470.
The main reason for the failure of chemotherapy therapies based on 5-Fluorouracil (5-Fu) is the development of resistance to 5-Fu in cancer patients, particularly those with colorectal cancer. Tissue inhibitor of metalloproteinases 2 (TIMP-2) has been shown to be associated with colorectal cancer (CRC), but its correlation with 5-Fu resistance in colorectal cancer has not been thoroughly studied. We screen the expression of different cytokines through Cytokine array. CCK-8 assay was conducted to evaluate the IC of 5-Fu and cell proliferation. ELISA and RT-qPCR were performed to detect TIMP-2 expression levels in cells and patient serum. Western blotting was utilised to analyse the differences in the expression of proteins related to signalling pathways in cells. Through cytokine array screening, we found that the expression of TIMP-2 was significantly increased in CRC drug-resistant cell lines. In addition, the expression of TIMP-2 in the serum of patients with CRC resistance to 5-Fu was significantly increased. Subsequent mechanistic experiments showed that TIMP-2 regulated the resistance of CRC cells to 5-Futhrough the JAK-STAT signalling pathway. Moreover, anti-TIMP-2 antibody or small molecule drug LY2784544 targeting the JAK-STAT signalling pathway can effectively reverse the resistance of CRC cells to 5-Fu. It is exactly TIMP-2 that mediates the resistance of CRC to 5-Fu through the JAK-STAT signalling pathway. Targeting drugs for TIMP-2 or the JAK-STAT signalling pathway are expected to be opportunities to reverse 5-Fu resistance in CRC.
基于5-氟尿嘧啶（5-Fu）的化疗疗法失败的主要原因是癌症患者，尤其是结直肠癌患者对5-Fu产生耐药性。金属蛋白酶组织抑制剂2（TIMP-2）已被证明与结直肠癌（CRC）相关，但其与结直肠癌中5-Fu耐药性的相关性尚未得到充分研究。我们通过细胞因子阵列筛选不同细胞因子的表达。进行CCK-8测定以评估5-Fu的半数抑制浓度（IC）和细胞增殖。采用酶联免疫吸附测定（ELISA）和逆转录定量聚合酶链反应（RT-qPCR）检测细胞和患者血清中TIMP-2的表达水平。利用蛋白质免疫印迹法分析细胞中与信号通路相关蛋白质表达的差异。通过细胞因子阵列筛选，我们发现TIMP-2在CRC耐药细胞系中的表达显著增加。此外，对5-Fu耐药的CRC患者血清中TIMP-2的表达也显著增加。随后的机制实验表明，TIMP-2通过JAK-STAT信号通路调节CRC细胞对5-Fu的耐药性。此外，抗TIMP-2抗体或靶向JAK-STAT信号通路的小分子药物LY2784544可有效逆转CRC细胞对5-Fu的耐药性。正是TIMP-2通过JAK-STAT信号通路介导了CRC对5-Fu的耐药性。针对TIMP-2或JAK-STAT信号通路的靶向药物有望成为逆转CRC中5-Fu耐药性的契机。
