晚期胆道癌（BTC）患者预后差，急需新的治疗方法。吉西他滨（gemcitabine）是BTC的标准治疗药物，它能引起DNA损伤，然而，癌细胞修复DNA的能力削弱了其疗效。为了提高吉西他滨的疗效，我们将其与MK1775（Wee1抑制剂）联合使用，后者可以阻止G2/M检查点的激活。我们对BTC细胞系分别用吉西他滨独立应用和与MK1775联合应用进行治疗，以确定其在BTC中的治疗潜力。结果显示，当与MK1775联合应用时，吉西他滨对四种BTC细胞系的生长抑制和诱导凋亡作用较单独应用时更明显。该联合治疗的效果在p53野生型和p53突变型细胞中均观察到，并且野生型p53的敲低对其效果没有影响。联合治疗增加了凋亡细胞的百分比，降低了合成DNA的细胞百分比，表明其导致了在S期中受损DNA的细胞积累和可能死亡。当细胞在细胞分裂停滞剂苏州诺使用下处于有丝分裂状态时，该联合治疗并未诱导凋亡。在移植瘤小鼠模型中，吉西他滨与MK1775结合（但其中任何一个药物单独使用不行）抑制了接种BTC细胞产生的肿瘤的生长。我们的结果表明，MK1775无论p53基因的状态如何，都能极大增强吉西他滨对BTC细胞的细胞毒性。我们认为，该联合治疗引发DNA损伤反应，并导致细胞凋亡。我们的临床前研究为将吉西他滨与MK1775联合用于BTC患者的未来临床试验奠定了基础。版权© 2023 The Authors. Elsevier Masson SAS发表并保留所有权利。

Patients with advanced biliary tract cancer (BTC) have a poor prognosis, and novel treatments are needed. Gemcitabine, the standard of care for BTC, induces DNA damage; however, the ability of cancer cells to repair DNA dampens its effects. To improve the efficacy of gemcitabine, we combined it with MK1775, a Wee1 inhibitor that prevents activation of the G2/M checkpoint. BTC cell lines were treated with gemcitabine only or in combination with MK1775 to determine the therapeutic potential of BTC. Gemcitabine inhibited the growth and induced the apoptosis of four BTC cell lines to a greater extent when added with MK1775 than when added alone. The effects of the combination treatment were observed in both p53 wild-type and p53 mutant cell lines and were unaffected by knockdown of wild-type p53. The combination treatment increased the percentage of apoptotic cells and decreased the percentage of cells synthesizing DNA, suggesting that it caused DNA-damaged cells to accumulate and possibly die in S phase. It did not induce apoptosis when cells were arrested in mitosis using nocodazole. In a xenograft mouse model, gemcitabine plus MK1775 (but not either alone) inhibited the growth of tumors generated from inoculated BTC cells. Our results show that MK1775 highly enhances gemcitabine cytotoxicity in BTC regardless of p53 status. We suggest that the combination treatment elicits a DNA damage response and consequent apoptosis. Our preclinical study provides a basis for future clinical trials of gemcitabine plus MK1775 in patients with BTC.Copyright © 2023 The Authors. Published by Elsevier Masson SAS.. All rights reserved.