类泛素化激活酶抑制剂PEV通过细胞衰老提高结肠癌细胞对5-氟尿嘧啶的敏感性  

Neddylation Activating Enzyme Inhibitor PEV Enhances the Sensitivity of Colorectal Cancer Cells to 5-Fluorouracil through Cell Senescence

作　　者：花雨昕 倪梦珍 孙晴晴 翟潇晴 潘巍巍 HUA Yuxin;NI Mengzhen;SUN Qingqing;ZHAI Xiaoqing;PAN Weiwei(School of Basic Medical Science,Zhejiang Chinese Medical University,Hangzhou 310000,China;Department of Cell Biology,College of Medicine,Jiaxing University,Jiaxing 314001,China)

机构地区：[1]浙江中医药大学,基础医学院,杭州310000 [2]嘉兴大学医学院,细胞生物学教研室,嘉兴314001

出　　处：《中国细胞生物学学报》2024年第6期1100-1111,共12页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:31871402);浙江省自然科学基金(批准号:LY21H160047、LGD21H160003、LQ23C070001);“创新嘉兴•优才支持计划”先锋型创新团队资助的课题。

摘　　要：细胞衰老与肿瘤细胞增殖减缓密切相关,细胞衰老抑制了许多癌基因的激活,因此诱导细胞衰老是现阶段一种干扰肿瘤细胞增殖过程的潜在策略,或许可以成为治疗肿瘤的有效途径。PEV(Pevonedistat)又称MLN4924,是NEDD8激活酶(NAE)抑制剂,可以通过阻断Cullin类泛素化使Cullin-RING E3泛素连接酶(CRL)失活。PEV可以抑制肿瘤细胞增殖,促进细胞凋亡、细胞衰老。5-氟尿嘧啶(5-FU)是结肠癌治疗中的一线化疗药物。PEV是否可以通过促进细胞衰老增强化疗药物5-FU的敏感性,提高结肠癌的治疗效果尚未可知。该研究对结肠癌细胞HCT116、HT29分别进行PEV单药处理、5-FU单药处理以及PEV与5-FU联合处理,通过β-半乳糖苷酶染色、Western blot、细胞3天连续计数、CCK-8实验、裸鼠皮下成瘤实验、免疫组化等实验对结肠癌细胞的衰老与增殖情况进行探究。结果提示,与溶媒组相比,PEV处理组与PEV、5-FU联合处理组半乳糖苷酶活性增加,P21、P27、PAI-1、P62等衰老相关蛋白表达水平增加以及细胞核形态标志物LaminB1缺失。同时与PEV单药处理组相比,PEV、5-FU联合处理组细胞衰老程度增加,对细胞增殖的抑制效果更为显著。该研究揭示类泛素化激活酶抑制剂PEV促进细胞衰老提高结肠癌对5-FU的敏感性,从而加剧细胞衰老、抑制细胞增殖。Cell senescence is closely related to the slowing down of tumor cell proliferation,and cell senescence inhibits the activation of many oncogenes.Therefore,inducing cell senescence is a potential strategy to interfere with the important process of tumor cells proliferation at this stage,and may be an effective way to treat tumors.PEV(Pevonedistat),also known as MLN4924,is a NAE(NEDD8-activating enzyme)inhibitor that can inactivate CRL by blocking Cullin neddylation.PEV can inhibit the proliferation of tumor cells,promote cell apoptosis and senescence.5-FU(5-fluorouracil)is a first-line chemotherapy agent in the treatment of colorectal cancer.However,it is not known whether PEV can enhance the sensitivity of chemotherapy drug 5-FU and improve the therapeutic effect of colorectal cancer by promoting cell senescence.In this study,colorectal cancer cells HCT116 and HT29 were treated with PEV alone,5-FU alone and PEV combined with 5-FU respectively.Throughβ-galactosidase staining,Western blot,cell count for three days,CCK-8 test,mice and xenograft models,immunohistochemistry and other experiments,it was proved that PEV promoted cell senescence,the combination of PEV and 5-FU accelerated cell senescence and increased the sensitivity of colorectal cancer to 5-FU.The inhibitory effect on cell proliferation was more significant.The results showed that compared with the solvent group,the galactosidase activity,the expression of senescence-related proteins such as P21,P27,PAI-1,P62 and the loss of nuclear morphological marker LaminB1 were increased in PEV plus PEV and 5-FU groups.At the same time,compared with PEV single drug treatment group,PEV and 5-FU combined treatment group increased cell senescence and inhibited cell proliferation more significantly.This study revealed that NEDD8-activating enzyme inhibitor PEV promotes cell senescence and increases the sensitivity of colorectal cancer to 5-FU,which aggravates cell senescence and inhibits cell proliferation.

关 键 词：类泛素化 细胞衰老 结肠癌 化疗药物敏感性 

分 类 号：R735.35[医药卫生—肿瘤]