Ayaz Muhammad, Sadiq Abdul, Wadood Abdul, Junaid Muhammad, Ullah Farhat, Zaman Khan Nadir
Department of Pharmacy, University of Malakand, Chakdara, 18000 Dir (L), KPK, Pakistan.
Department of Pharmacy, University of Malakand, Chakdara, 18000 Dir (L), KPK, Pakistan; Department of Life Sciences & Chemistry, Faculty of Health, Jacobs University Bremen, Germany.
Steroids. 2019 Jan;141:30-35. doi: 10.1016/j.steroids.2018.11.005. Epub 2018 Nov 13.
Based on our previous studies on cytotoxic potentials of Polygonum hydropiper L, two steroidal compounds beta-sitosterol and stigmasterol were isolated from the most active fraction and were subjected to cell lines cytotoxicity. Isolated compounds were tested against HeLa, MCF-7 and NIH/3T3 cell lines following MTT assay. Furthermore, the compounds were also docked against tyrosine kinase enzyme to predict the binding mode of phytosterols in the active sites of the enzyme. Beta-sitosterol exhibited considerable cytotoxicity against NIH/3T3, HeLa and MCF-7 cell with 67.05 ± 2.08, 79.63 ± 2.34 and 71.50 ± 1.57% lethality respectively at 1 mg/ml concentration. Median inhibitory concentrations calculated from dose response curve against NIH/3T3, HeLa and MCF-7 cells were 440, 170 and 200 µg/ml respectively. Stigmasterol was more effective against MCF-7 and NIH/3T3 cells by killing 87.50 and 81.45% cancerous cells respectively at 1 mg/ml concentration. Stigmasterol showed 77.25% cyctotoxicity against HeLA cells at 1 mg/ml concentration in MTT assay. The IC values for HeLA, MCF-7 and NIH/3T3 cells were 170, 60 and 140 µg/ml respectively. In docking studies, the docking score for beta-sitosterol and stigmasterol were -7.266 and -4.89 respectively. The binding energies for beta-sitosterol and stigmasterol were -41.21 and -41.04 respectively. Such lower binding energies indicate that the compounds fit into the active site more strongly. Binding affinities for both compounds were -7.76 and -7.68 respectively. Both phytosterols possess significant anticancer potentials and can be effective in the prevention and treatment of several malignancies.
基于我们之前对水蓼细胞毒性潜力的研究，从活性最高的组分中分离出两种甾体化合物β-谷甾醇和豆甾醇，并对其进行细胞系细胞毒性实验。采用MTT法，将分离得到的化合物对HeLa、MCF-7和NIH/3T3细胞系进行测试。此外，还将这些化合物与酪氨酸激酶进行对接，以预测植物甾醇在该酶活性位点的结合模式。β-谷甾醇在1mg/ml浓度下对NIH/3T3、HeLa和MCF-7细胞表现出相当大的细胞毒性，致死率分别为67.05±2.08%、79.63±2.34%和71.50±1.57%。根据针对NIH/3T3、HeLa和MCF-7细胞的剂量反应曲线计算出的半数抑制浓度分别为440、170和200μg/ml。豆甾醇在1mg/ml浓度下对MCF-7和NIH/3T3细胞更有效，分别杀死87.50%和81.45%的癌细胞。在MTT实验中，豆甾醇在1mg/ml浓度下对HeLA细胞表现出77.25%的细胞毒性。HeLA、MCF-7和NIH/3T3细胞的IC值分别为170、60和140μg/ml。在对接研究中，β-谷甾醇和豆甾醇的对接分数分别为-7.266和-4.89。β-谷甾醇和豆甾醇的结合能分别为-41.21和-41.04。如此低的结合能表明这些化合物与活性位点的结合更强。两种化合物的结合亲和力分别为-7.76和-7.68。两种植物甾醇都具有显著的抗癌潜力，可有效预防和治疗多种恶性肿瘤。
