背景:阑尾癌(AC)粘蛋白的过量产生是加热腹膜内化疗(HIPEC)药物递送的障碍。菠萝蛋白酶是一种菠萝茎提取物,具有粘液溶解特性。我们在患者衍生的肿瘤类器官(PTO)模型和AC细胞系中探索了菠萝蛋白酶对粘液AC的治疗效果。
方法:PTO是从接受HIPEC细胞减灭术的AC患者获得的肿瘤标本中制作的。PTO接受菠萝蛋白酶HIPEC治疗,顺铂,和丝裂霉素C(MMC)在37°C和42°C下进行和不进行菠萝蛋白酶预处理。
结果:从2020年10月至2023年5月,从13例低级别(12/16,75%)和高级别AC(4/16,25%)的患者中收集了16个标本。与菠萝蛋白酶相比,菠萝蛋白酶与N-乙酰半胱氨酸(NAC)联合使用时的粘蛋白消耗作用最为显着(47%对10%,p=0.0009)或单独使用NAC(47%对12.8%,p=0.0027)。菠萝蛋白酶在60分钟时显示>31%的类器官活力降低(p<0.001),在48小时时显示>66%(p<0.0001)。菠萝蛋白酶预处理使顺铂和MMCHIPEC条件的细胞毒性增加了31.6%(p=0.0001)和35.5%(p=0.0001),分别。菠萝蛋白酶处理后,Ki67,CK20和MUC2表达降低;而caspase3/7活性增加,Bcl-2(p=0.009)和Bcl-xL(p=0.01)降低,提示诱导凋亡途径。此外,自噬蛋白LC3A/BI(p<0.03)和II(p<0.031)增加;而ATG7(p<0.01),ATG12(p<0.04),和Becline1(p<0.03),在菠萝蛋白酶处理的PTO中表达降低。
结论:菠萝蛋白酶显示出针对阑尾癌类器官的细胞毒性和粘液溶解活性。作为预处理剂,它增强了多种HIPEC方案的细胞毒性,可能通过程序性细胞死亡和自噬介导。
BACKGROUND: Appendiceal cancer (AC) excessive mucin production is a barrier to heated intraperitoneal chemotherapy (HIPEC) drug delivery. Bromelain is a pineapple stem extract with mucolytic properties. We explored bromelain treatment effects against mucinous AC in a patient-derived tumor organoid (PTO) model and an AC cell line.
METHODS: PTOs were fabricated from tumor specimens obtained from patients with AC undergoing cytoreductive surgery with HIPEC. PTOs underwent HIPEC treatment with bromelain, cisplatin, and mitomycin C (MMC) at 37 °C and 42 °C with and without bromelain pretreatment.
RESULTS: From October 2020 to May 2023, 16 specimens were collected from 13 patients with low-grade (12/16, 75%) and high-grade AC (4/16, 25%). The mucin-depleting effects of bromelain were most significant in combination with N-acetylcysteine (NAC) compared with bromelain (47% versus 10%, p = 0.0009) or NAC alone (47% versus 12.8%, p = 0.0027). Bromelain demonstrated > 31% organoid viability reduction at 60 min (p < 0.001) and > 66% in 48 h (p < 0.0001). Pretreatment with bromelain increased cytotoxicity of both cisplatin and MMC HIPEC conditions by 31.6% (p = 0.0001) and 35.5% (p = 0.0001), respectively. Ki67, CK20, and MUC2 expression decreased after bromelain treatment; while increased caspase 3/7 activity and decreased Bcl-2 (p = 0.009) and Bcl-xL (p = 0.01) suggest induction of apoptosis pathways. Furthermore, autophagy proteins LC3A/B I (p < 0.03) and II (p < 0.031) were increased; while ATG7 (p < 0.01), ATG 12 (p < 0.04), and Becline 1(p < 0.03), expression decreased in bromelain-treated PTOs.
CONCLUSIONS: Bromelain demonstrates cytotoxicity and mucolytic activity against appendiceal cancer organoids. As a pretreatment agent, it potentiates the cytotoxicity of multiple HIPEC regimens, potentially mediated through programmed cell death and autophagy.