Xie Ting, Wang Yunyun, Guo Ting, Yuan Chunhua
Department of Neurobiology, School of Basic Medical Sciences, Southern Medical University, Guangdong-Hong Kong-Macao Greater Bay Area Center for Brain Science and Brain-Inspired Intelligence, Guangzhou 510515, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2025 Jul 20;45(7):1460-1470. doi: 10.12122/j.issn.1673-4254.2025.07.12.
To evaluate the inhibitory effect of crude venom against proliferation of different cancer cells and identify the active components in the venom.
Different cancer cell lines were treated with different concentrations of venom for 48 h, and cell proliferation and the half-maximal inhibitory concentrations (IC) of the venom were assessed with CCK-8 assay. The apoptosis rate of breast cancer MCF7 cells following the treatment was analyzed with flow cytometry, and the changes in cellular caspase-8 and caspase-9 expressions were detected. The crude venom was separated into protein, peptide, and small-molecule compound fractions using gel filtration chromatography and high-performance liquid chromatography (HPLC). The protein and peptide components were identified using proteomics analysis, and small-molecule compounds were structurally characterized using nuclear magnetic resonance (NMR), mass spectrometry (MS), and HPLC.
The crude venom exhibited strong concentration-dependent inhibitory effects on proliferation of MCF7 cells and nasopharyngeal carcinoma SUNE1 and HONE1 cells (IC of 2.14±0.29, 1.57±0.14, and 2.85±0.15 µg/mL, respectively), with less potent inhibitory effects in gastric cancer HGC27 cells and colorectal cancer SW620 cells (IC of 3.02±0.27 and 3.02±0.28 µg/mL, respectively). The crude venom significantly promoted MCF7 cell apoptosis likely the caspase 8 signaling pathway. The protein fraction from the crude venom showed a weak inhibitory effect in MCF7 cells, whereas the peptide fraction exhibited a much stronger inhibitory effect (IC of 6.41±0.31 µg/mL). The peptides in the peptide fraction, with relative molecular mass around 10 000, were homologous to those found in venom. The small-molecule fraction consisted mainly of nucleotide metabolites without obvious inhibitory effects in MCF7 cells, but its combination with the peptide fraction showed significantly enhanced inhibitory activity. C The inhibitory effects of venom, which vary significantly across different cancer cell lines, are attributed primarily to its peptide components, which may act synergistically with the nucleotide metabolites.
评估粗毒对不同癌细胞增殖的抑制作用，并鉴定粗毒中的活性成分。
用不同浓度的粗毒处理不同癌细胞系48小时，采用CCK-8法评估细胞增殖及粗毒的半数抑制浓度（IC）。用流式细胞术分析处理后乳腺癌MCF7细胞的凋亡率，并检测细胞中半胱天冬酶-8和半胱天冬酶-9表达的变化。利用凝胶过滤色谱法和高效液相色谱法（HPLC）将粗毒分离为蛋白质、肽和小分子化合物组分。采用蛋白质组学分析鉴定蛋白质和肽成分，利用核磁共振（NMR）、质谱（MS）和HPLC对小分子化合物进行结构表征。
粗毒对MCF7细胞、鼻咽癌SUNE1和HONE1细胞的增殖表现出强烈的浓度依赖性抑制作用（IC分别为2.14±0.29、1.57±0.14和2.85±0.15μg/mL），对胃癌HGC27细胞和结直肠癌SW620细胞的抑制作用较弱（IC分别为3.02±0.27和3.02±0.28μg/mL）。粗毒可能通过半胱天冬酶8信号通路显著促进MCF7细胞凋亡。粗毒中的蛋白质组分对MCF7细胞的抑制作用较弱，而肽组分表现出更强的抑制作用（IC为6.41±0.31μg/mL）。肽组分中相对分子质量约为10000的肽与粗毒中的肽同源。小分子组分主要由核苷酸代谢产物组成，对MCF7细胞无明显抑制作用，但其与肽组分联合时显示出显著增强的抑制活性。粗毒对不同癌细胞系的抑制作用差异显著，主要归因于其肽成分，肽成分可能与核苷酸代谢产物协同发挥作用。
