PDF(Pubmed)
Abstract：
The high recurrence rate of cervical cancer is a leading cause of cancer deaths in women. 5-Fluorouracil (5-FU) is an antitumor drug used to treat many types of cancer, but its diminishing effectiveness and side effects limit its use. Norcantharidin (NCTD), a demethylated derivative of cantharidin, exhibits various biological activities. Here, we investigated whether NCTD could potentiate 5-FU to induce cervical cancer cell death. To assess the cell viability and synergistic effects of the drugs, cell counting kit-8 and colony formation assays were performed using HR-HPV-positive cervical cancer cell lines. Annexin V-FITC/PI staining and TUNEL assays were performed to confirm the induction of apoptosis. The synergistic effect of NCTD on the antitumor activity of 5-FU was analyzed using network pharmacology, molecular docking, and molecular dynamics simulations. Apoptosis-related proteins were examined using immunoblotting. The combination of NCTD and 5-FU was synergistic in cervical cancer cell lines. Network pharmacological analysis identified 10 common targets of NCTD and 5-FU for cervical cancer treatment. Molecular docking showed the strong binding affinity of both compounds with CA12, CASP9, and PTGS1. Molecular dynamics simulations showed that the complex system of both drugs with caspase-9 could be in a stable state. NCTD enhanced 5-FU-mediated cytotoxicity by activating apoptosis-related proteins. NCTD acts synergistically with 5-FU to inhibit cervical cancer cell proliferation. NCTD enhances 5-FU-induced apoptosis in cervical cancer cell lines via the caspase-dependent pathway.
摘要：
宫颈癌的高复发率是女性癌症死亡的主要原因。5-氟尿嘧啶（5-FU）是一种用于治疗多种类型癌症的抗肿瘤药物，但是它的功效和副作用的减少限制了它的使用。Norcantharidin(NCTD),斑疹素的去甲基化衍生物,具有多种生物活性。这里,我们研究了NCTD是否可以增强5-FU诱导宫颈癌细胞死亡。为了评估药物的细胞活力和协同作用，使用HR-HPV阳性宫颈癌细胞系进行细胞计数试剂盒-8和集落形成测定.进行膜联蛋白V-FITC/PI染色和TUNEL测定以确认细胞凋亡的诱导。网络药理学分析了NCTD对5-FU抗肿瘤活性的协同作用,分子对接,和分子动力学模拟。使用免疫印迹检查凋亡相关蛋白。NCTD和5-FU的组合在宫颈癌细胞系中是协同的。网络药理学分析确定了NCTD和5-FU治疗宫颈癌的10个常见靶标。分子对接显示两种化合物与CA12、CASP9和PTGS1的强结合亲和力。分子动力学模拟表明，两种药物与caspase-9的复合系统均处于稳定状态。NCTD通过激活凋亡相关蛋白增强5-FU介导的细胞毒性。NCTD与5-FU协同作用以抑制宫颈癌细胞增殖。NCTD通过caspase依赖性途径增强5-FU诱导的宫颈癌细胞系凋亡。
