Graduate Institute of Clinical Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.
Department of Surgery, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.
PLoS One. 2023 Jun 1;18(6):e0285970. doi: 10.1371/journal.pone.0285970. eCollection 2023.
Colorectal cancer (CRC) is one of the most lethal cancers worldwide. Long-term survival is not achieved in metastatic CRC despite the current multidisciplinary therapies. Bromelain, a compound extracted from the pineapple plant, has multiple functions and anticancer properties. Previously, bromelain has been chromatographically separated into four fractions. Fraction 3 (F3) exhibits the highest proteolytic activity. The anticancer effects of F3 bromelain in CRC cells is unknown.
In vitro cytotoxicity was verified through a sulforhodamine B assay. Apoptosis in CRC cells induced by unfractionated or F3 bromelain was examined using Annexin V-FITC/PI staining and Western blot analysis. ROS status, autophagy and lysosome formation were determined by specific detection kit.
The cytotoxicity of F3 bromelain in CRC cells was found to be comparable to that of unfractionated bromelain. F3 bromelain induces caspase-dependent apoptosis in CRC cells. Treatment with unfractionated or F3 bromelain increased superoxide and oxidative stress levels and autophagy and lysosome formation. ATG5/12 and beclin-1 were upregulated, and the conversion of LC3B-I to LC3B-II was increased significantly by treatment with F3 bromelain. Treated CQ, autophagy inhibitor, with unfractionated or F3 bromelain enhances the cytotoxic effects. Finally, the combination of unfractionated and F3 bromelain with a routine chemotherapeutic agent (5-fluourouracil, irinotecan, or oxaliplatin) resulted in synergistically higher cytotoxic potency in CRC cells.
Unfractionated and F3 bromelain inhibits CRC cell proliferation in vitro, and the cytotoxic effects of unfractionated bromelain are equivalent to F3 bromelain. F3 bromelain may be a potential and potent drug for clinical use due to its anticancer efficacy and high synergistic cytotoxicity when combined with a routine chemotherapeutic agent for CRC.
结直肠癌（CRC）是全球最致命的癌症之一。尽管目前采用了多学科治疗方法，但转移性 CRC 仍无法实现长期生存。菠萝蛋白酶是从菠萝植物中提取的一种化合物，具有多种功能和抗癌特性。此前，菠萝蛋白酶已通过色谱分离成四个馏分。馏分 3（F3）表现出最高的蛋白水解活性。F3 菠萝蛋白酶在 CRC 细胞中的抗癌作用尚不清楚。
通过磺酰罗丹明 B 测定法验证体外细胞毒性。通过 Annexin V-FITC/PI 染色和 Western blot 分析检测未分级或 F3 菠萝蛋白酶诱导的 CRC 细胞凋亡。通过特定检测试剂盒确定 ROS 状态、自噬和溶酶体形成。
发现 F3 菠萝蛋白酶在 CRC 细胞中的细胞毒性与未分级菠萝蛋白酶相当。F3 菠萝蛋白酶诱导 CRC 细胞中 caspase 依赖性细胞凋亡。用未分级或 F3 菠萝蛋白酶处理会增加超氧化物和氧化应激水平以及自噬和溶酶体形成。ATG5/12 和 beclin-1 上调，F3 菠萝蛋白酶处理后 LC3B-I 向 LC3B-II 的转化明显增加。用自噬抑制剂 CQ 处理未分级或 F3 菠萝蛋白酶可增强细胞毒性作用。最后，将未分级和 F3 菠萝蛋白酶与常规化疗药物（5-氟尿嘧啶、伊立替康或奥沙利铂）联合使用可使 CRC 细胞的协同细胞毒性作用显著提高。
未分级和 F3 菠萝蛋白酶可抑制 CRC 细胞在体外的增殖，未分级菠萝蛋白酶的细胞毒性作用与 F3 菠萝蛋白酶相当。F3 菠萝蛋白酶可能是一种有潜力的药物，因其抗癌作用以及与常规化疗药物联合使用时具有高协同细胞毒性作用，可用于临床。
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