Jeffrey Cheah School of Medicine and Health Sciences, Monash University Malaysia, Bandar Sunway, Selangor Darul Ehsan, Malaysia.
Kuliyyah of Pharmacy, International Islamic University Malaysia, Kuantan, Pahang Darul Makmur, Malaysia.
Basic Clin Pharmacol Toxicol. 2018 Nov;123(5):577-588. doi: 10.1111/bcpt.13060. Epub 2018 Jul 25.
The aim of this study was to investigate the cytotoxic, antiproliferative activity and the induction of apoptosis by L-amino acid oxidase isolated from Calloselasma rhodostoma crude venom (CR-LAAO) on human colon cancer cells. CR-LAAO was purified using three chromatographic steps: molecular exclusion using G-50 gel filtration resin, ion-exchange by MonoQ column and desalted on a G25 column. The purity and identity of the isolated CR-LAAO was confirmed by SDS-PAGE and LC-MS/MS. CR-LAAO demonstrated time- and dose-dependent cytotoxic activity on SW480 (primary human colon cancer cells) and SW620 (metastatic human colon cancer cells) with an EC values of 6 μg/ml and 7 μg/ml at 48 hr, respectively. Quantification of apoptotic cells based on morphological features demonstrated significant increase in apoptotic cell population in both SW480 and SW620 cells which peaked at 48 hr. Significant increase in caspase-3 activity and reduction in Bcl-2 levels were demonstrated following CR-LAAO treatment. These data provide evidence on the potential anticancer activity of CR-LAAO from the venom of C. rhodostoma for therapeutic intervention of human colon cancer.
本研究旨在探讨从尖吻蝮粗毒中分离得到的 L-氨基酸氧化酶（CR-LAAO）对人结肠癌细胞的细胞毒性、抗增殖活性和诱导细胞凋亡的作用。CR-LAAO 通过三种色谱步骤进行纯化：使用 G-50 凝胶过滤树脂进行分子排阻、使用 MonoQ 柱进行离子交换以及在 G25 柱上脱盐。分离得到的 CR-LAAO 的纯度和身份通过 SDS-PAGE 和 LC-MS/MS 得到确认。CR-LAAO 对 SW480（原发性人结肠癌细胞）和 SW620（转移性人结肠癌细胞）表现出时间和剂量依赖性的细胞毒性作用，在 48 小时时 EC 值分别为 6μg/ml 和 7μg/ml。基于形态特征的凋亡细胞定量显示，在 SW480 和 SW620 细胞中，凋亡细胞群体显著增加，在 48 小时时达到峰值。CR-LAAO 处理后， caspase-3 活性显著增加，Bcl-2 水平降低。这些数据为尖吻蝮蛇毒中的 CR-LAAO 具有治疗人类结肠癌的潜在抗癌活性提供了证据。
