El-Naggar Sabry Ali, Basyony Mohamed Aboulfotouh, El-Wahsh Hany M, El-Feki Seham Mohamed, Kandyel Ramadan Mahmoud
Zoology Department, Faculty of Science, Tanta University, Al-Geish St., Tanta, Gharbia 31527, Egypt.
Marine Biology Department, Faculty of Marine Sciences, King Abdulaziz University, Jeddah 21589, Saudi Arabia.
Trop Life Sci Res. 2024 Jul;35(2):65-85. doi: 10.21315/tlsr2024.35.2.4. Epub 2024 Jul 31.
This study aims to determine the biochemical compositions and the antitumour effect of the parotoid gland secretions (PGS) of the Egyptian toad (). The total protein, lipid, carbohydrate contents, total antioxidant capacity (TAC), the median inhibitory concentration (IC) of 2,2-diphenyl-1-picrylhydrazyl (DPPH), sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) profile, amino acid analysis, gas chromatography-mass spectrometry (GC-MS) analysis and minerals were determined in PGS. The antitumour effect of PGS against human hepatocellular carcinoma (HepG-2), breast adenocarcinoma (MCF-7) and normal lung fibroblast (WI-38) cell lines were determined. The total protein, lipid and carbohydrate contents of PGS were 250 ± 15 mg/g D.W, 33 ± 3.2 mg/g D.W and 5 ± 0.65 mg/g D.W, respectively, while its TAC was 16.56 ± 0.12 mg/g D.W and the IC of DPPH was 51.95 ± 2.95 mg/mL. Six protein bands varied between 10 and 50 kDa were found in PGS. Among amino acid profile, arginine showed the highest content in PGS. GC-MS analysis showed that 11-octadecenoic acid methyl ester was the highest concentrations in PGS. The half-maximal inhibitory concentrations (IC) of PGS against HepG-2, MCF-7 and WI-38 cells were 131.82 ± 6.14, 189.71 ± 8.95 and 685.65 ± 33.1 μg/mL, respectively. study showed that treatment of HepG-2 and MCF-7 cells with PGS increased the percentages of early and late apoptotic. While the percentages of early and late apoptotic WI-38 cells after treatment with PGS were 2.1% and 3.7%. Cell cycle analysis showed that PGS treatment arrested HepG-2 and WI-38 in S-phase, while arrested MCF-7 cells in G2/M phase. The present study concluded that PGS has a potent antioxidant activity with antitumour effect against HepG-2 and MCF-7 cells.
本研究旨在确定埃及蟾蜍腮腺分泌物（PGS）的生化成分及其抗肿瘤作用。测定了PGS中的总蛋白、脂质、碳水化合物含量、总抗氧化能力（TAC）、2,2-二苯基-1-苦基肼（DPPH）的半数抑制浓度（IC）、十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）图谱、氨基酸分析、气相色谱-质谱联用（GC-MS）分析以及矿物质含量。测定了PGS对人肝癌细胞（HepG-2）、乳腺腺癌细胞（MCF-7）和正常肺成纤维细胞（WI-38）细胞系的抗肿瘤作用。PGS的总蛋白、脂质和碳水化合物含量分别为250±15mg/g干重、33±3.2mg/g干重和5±0.65mg/g干重，其TAC为16.56±0.12mg/g干重，DPPH的IC为51.95±2.95mg/mL。在PGS中发现了6条分子量在10至50kDa之间的蛋白条带。在氨基酸谱中，精氨酸在PGS中的含量最高。GC-MS分析表明，11-十八碳烯酸甲酯在PGS中的浓度最高。PGS对HepG-2、MCF-7和WI-38细胞的半数最大抑制浓度（IC）分别为131.82±6.14、189.71±8.95和685.65±33.1μg/mL。研究表明，用PGS处理HepG-2和MCF-7细胞可增加早期和晚期凋亡的百分比。而用PGS处理后，WI-38细胞的早期和晚期凋亡百分比分别为2.1%和3.7%。细胞周期分析表明，PGS处理使HepG-2和WI-38细胞停滞在S期，使MCF-7细胞停滞在G2/M期。本研究得出结论，PGS具有强大的抗氧化活性，对HepG-2和MCF-7细胞具有抗肿瘤作用。
