背景:喜树碱(CPT),一种具有抗肿瘤特性的五环生物碱,来自喜树。拓扑替康和伊立替康(CPT衍生物)在25年前首次被食品和药物管理局批准用于癌症治疗,并且仍然是今天的关键抗癌药物。然而,它们的使用通常受到临床毒性的限制。尽管进行了广泛的发展努力,许多这些衍生物在临床上没有成功,特别是它们对胰腺癌的有效性仍然很低。
目的:本研究旨在评估我们实验室合成的CPT衍生物FLQY2的治疗活性,对抗胰腺癌,将其功效和作用机制与已建立的临床药物进行比较。
方法:使用MTT测定法评估FLQY2对癌细胞的细胞毒性作用。采用患者来源的类器官(PDO)模型来比较FLQY2对各种癌症中现有临床药物的敏感性。通过流式细胞术检测FLQY2对MiaPaca-2胰腺癌细胞凋亡和细胞周期停滞的影响。进行转录组和蛋白质组分析以探索FLQY2抗肿瘤活性的潜在机制。使用蛋白质印迹法确定由FLQY2调节的蛋白质的水平。此外,在胰腺癌异种移植模型中评估了FLQY2的体内抗肿瘤功效.
结果:FLQY2表现出(1)有效的细胞毒性;(2)与目前的临床药物如吉西他滨相比,在PDO模型中具有优异的肿瘤抑制活性,5-氟尿嘧啶,顺铂,紫杉醇,ivosidenib,infinitinib,和lenvatinib;(3)在胰腺癌异种移植模型中,肿瘤抑制作用明显高于紫杉醇脂质体;(4)强大的抗肿瘤作用,与TOPI和PDK1/AKT/mTOR信号通路的抑制密切相关。体外研究表明,FLQY2抑制细胞增殖,菌落形成,诱导细胞凋亡,并在纳摩尔浓度下引起细胞周期停滞。此外,FLQY2和吉西他滨的组合具有显著的抑制作用和协同作用。
结论:该研究证实了拓扑异构酶I和PDK1/AKT/mTOR通路参与介导FLQY2治疗MiaPaca-2胰腺癌的抗肿瘤活性。因此,FLQY2具有作为胰腺癌患者的新型治疗选择的潜力。
BACKGROUND: Camptothecin (CPT), a pentacyclic alkaloid with antitumor properties, is derived from the Camptotheca acuminata. Topotecan and irinotecan (CPT derivatives) were first approved by the Food and Drug Administration for cancer treatment over 25 years ago and remain key anticancer drugs today. However, their use is often limited by clinical toxicity. Despite extensive development efforts, many of these derivatives have not succeeded clinically, particularly in their effectiveness against pancreatic cancer which remains modest.
OBJECTIVE: This study aimed to evaluate the therapeutic activity of FLQY2, a CPT derivative synthesized in our laboratory, against pancreatic cancer, comparing its efficacy and mechanism of action with those of established clinical drugs.
METHODS: The cytotoxic effects of FLQY2 on cancer cells were assessed using an MTT assay. Patient-derived organoid (PDO) models were employed to compare the sensitivity of FLQY2 to existing clinical drugs across various cancers. The impact of FLQY2 on apoptosis and cell cycle arrest in Mia Paca-2 pancreatic cancer cells was examined through flow cytometry. Transcriptomic and proteomic analyses were conducted to explore the underlying mechanisms of FLQY2\'s antitumor activity. Western blotting was used to determine the levels of proteins regulated by FLQY2. Additionally, the antitumor efficacy of FLQY2 in vivo was evaluated in a pancreatic cancer xenograft model.
RESULTS: FLQY2 demonstrated (1) potent cytotoxicity; (2) superior tumor-suppressive activity in PDO models compared to current clinical drugs such as gemcitabine, 5-fluorouracil, cisplatin, paclitaxel, ivosidenib, infinitinib, and lenvatinib; (3) significantly greater tumor inhibition than paclitaxel liposomes in a pancreatic cancer xenograft model; (4) robust antitumor effects, closely associated with the inhibition of the TOP I and PDK1/AKT/mTOR signaling pathways. In vitro studies revealed that FLQY2 inhibited cell proliferation, colony formation, induced apoptosis, and caused cell cycle arrest at nanomolar concentrations. Furthermore, the combination of FLQY2 and gemcitabine exhibited significant inhibitory and synergistic effects.
CONCLUSIONS: The study confirmed the involvement of topoisomerase I and the PDK1/AKT/mTOR pathways in mediating the antitumor activity of FLQY2 in treating Mia Paca-2 pancreatic cancer. Therefore, FLQY2 has potential as a novel therapeutic option for patients with pancreatic cancer.