Institute of Medical Science Technology, Universiti Kuala Lumpur, 43000 Kajang, Selangor, Malaysia.
Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
Trop Biomed. 2024 Jun 1;41(2):160-165. doi: 10.47665/tb.41.2.005.
Helminth parasites are a group of complex metazoans from various taxonomic families. Excretory secretory (ES) by-products, secreted by living parasites from the surface, appeared to modulate the host immunological response towards helminth infection. This study aims to investigate the effect of ES antigen from helminth parasite on colorectal cell viability. Worm were cultured in phosphate-buffered saline (PBS x1) at 37°C for 24 hours after being rinsed in sterile PBS. Using a mortar and pestle, the worm was crushed vigorously using PBS. The obtained excretory secretory (ES) antigens were extracted and filtered using a 0.22 µM filter and stored at -20°C for further assay. For LCMS, 100 µl of the extract was analysed using Agilent ZORBAX Eclipse Plus C18 Rapid Resolution HT. The extraction of ES antigen (10 µg/ml and 20 µg/ml) was used for cell viability studies using CRC cell line HCT 116. Cell viability and MTT assay were conducted as per the protocol mentioned in the MTT kit. The liquid chromatography and mass spectrometry (LCMS) data indicated that the ES antigen contained metabolic compounds, namely fatty acid, amino alcohol, indoles, sterols, glycosides, and sphingoids. For the Ascaris lumbricoides LCMS analyses, around 405 metabolic peaks were detected. Out of which, 58 were detected via the database were identified, while several compounds detected have anticancer properties. The MTT assay indicated that after 24 hours and 48 hours of exposure, all treated cells showed a decrease in cell viability compared to the control group. The preliminary studies demonstrated that the ES antigen from Ascaris lumbricoides has some ability to decrease the cell viability of the HCT116 CRC cell line. Further studies are needed to examine the cell cycle arrest and apoptosis effect of the ES antigen towards the CRC cell line.
寄生虫是一组来自不同分类家族的复杂后生动物。寄生虫从表面分泌的排泄分泌（ES）副产物似乎调节了宿主对寄生虫感染的免疫反应。本研究旨在研究寄生虫 ES 抗原对结直肠细胞活力的影响。用磷酸盐缓冲盐水（PBS x1）冲洗后，将虫体在 37°C 下培养 24 小时。用研钵和杵将虫体剧烈压碎，用 PBS 冲洗。提取获得的排泄分泌（ES）抗原，用 0.22µM 过滤器过滤，并储存在-20°C 以备进一步检测。对于 LCMS，用 Agilent ZORBAX Eclipse Plus C18 Rapid Resolution HT 分析 100µl 提取物。使用 CRC 细胞系 HCT 116 进行细胞活力研究，使用 ES 抗原（10µg/ml 和 20µg/ml）进行提取。按照 MTT 试剂盒中的方案进行细胞活力和 MTT 测定。液相色谱和质谱（LCMS）数据表明 ES 抗原含有代谢化合物，如脂肪酸、氨基酸醇、吲哚、甾醇、糖苷和神经酰胺。对于蛔虫的 LCMS 分析，检测到约 405 个代谢峰。其中，通过数据库检测到 58 个，而检测到的几种化合物具有抗癌特性。MTT 测定表明，与对照组相比，所有处理细胞在暴露 24 小时和 48 小时后细胞活力均下降。初步研究表明，蛔虫 ES 抗原具有降低 HCT116 CRC 细胞系细胞活力的能力。需要进一步研究 ES 抗原对 CRC 细胞系的细胞周期停滞和凋亡作用。
