Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, China.
Department of Surgery, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, Jinhua 321000, China.
ACS Chem Biol. 2024 Nov 15;19(11):2336-2344. doi: 10.1021/acschembio.4c00446. Epub 2024 Oct 28.
Gemcitabine-based monotherapy or combination therapy has become the standard treatment for locally advanced and metastatic pancreatic cancer. However, the emergence of resistance within weeks of treatment severely compromises therapeutic efficacy. The intricate biological process of gemcitabine resistance in pancreatic cancer presents a complex challenge, as the underlying mechanisms remain unclear. Identifying the target protein of gemcitabine is crucial for studying its drug-resistance mechanism. An activity-based probe is a powerful tool for studying drug target proteins, but the current lack of activity-based gemcitabine probes with robust biological activity hinders research on gemcitabine. In this study, we developed three active probes based on gemcitabine, among which demonstrated excellent stability and labeling efficacy. We utilized in conjunction with chemical proteomics to identify intracellular target proteins. We identified 79 proteins that interact with gemcitabine, most of which were previously unknown and represented various functional classes. Additionally, we validated the increased expression of IFIT3 and MARCKS in drug-resistant cells, along with the activation of the NF-κB signaling pathway. These findings substantially contribute to our comprehension of gemcitabine's target proteins and further our understanding of the mechanisms driving gemcitabine resistance in pancreatic cancer cells.
基于吉西他滨的单药或联合治疗已成为局部晚期和转移性胰腺癌的标准治疗方法。然而，在治疗开始后的数周内出现的耐药性严重影响了治疗效果。胰腺癌中吉西他滨耐药性的复杂生物学过程是一个复杂的挑战，因为其潜在机制尚不清楚。鉴定吉西他滨的靶蛋白对于研究其耐药机制至关重要。基于活性的探针是研究药物靶蛋白的有力工具，但目前缺乏具有强大生物学活性的基于活性的吉西他滨探针，这阻碍了对吉西他滨的研究。在这项研究中，我们基于吉西他滨开发了三种活性探针，其中 表现出优异的稳定性和标记效果。我们利用 结合化学蛋白质组学来鉴定细胞内的靶蛋白。我们鉴定出了 79 种与吉西他滨相互作用的蛋白质，其中大多数是以前未知的，代表了各种功能类别。此外，我们验证了耐药细胞中 IFIT3 和 MARCKS 的表达增加，以及 NF-κB 信号通路的激活。这些发现极大地促进了我们对吉西他滨靶蛋白的理解，并进一步深入了解了驱动胰腺癌细胞中吉西他滨耐药性的机制。
