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.
Abstract
Cell senescence is closely related to the slowing down of tumor cell proliferation,and cell se-nescence 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 im-prove 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,immuno-histochemistry 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 ef-fect on cell proliferation was more significant.The results showed that compared with the solvent group,the galac-tosidase 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 in-hibited cell proliferation more significantly.This study revealed that NEDD8-activating enzyme inhibitor PEV pro-motes cell senescence and increases the sensitivity of colorectal cancer to 5-FU,which aggravates cell senescence and inhibits cell proliferation.
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