Department of Microbiology, Colleges of Basic Medical Sciences, Dalian Medical University , 9 Western Section, Lvshun South Street, Lvshunkou District, Dalian 116044, Liaoning Province, P.R. China.
J Agric Food Chem. 2014 Jun 11;62(23):5316-20. doi: 10.1021/jf500316f. Epub 2014 May 29.
We purified and isolated a novel protein (LFP(91-3)A2) with antitumor effect from Lentinula edodes C(91-3) liquid mycelial culture supernatant. LFP(91-3)A2 was purified by (NH4)2SO4 precipitation, ion-exchange chromatography (DEAE-cellulose) and gel filtration chromatography (Sephacryl S-200HR). SDS-PAGE and MALDI-TOF/MS analysis Mascot search showed LFP(91-3)A2 is a new protein with apparent molecular weight of 26 kDa. The effect on tumor cell proliferation was assessed by using MTT assay in vitro, and the LFP(91-3)A2 reduced tumor cell growth obviously in a dose dependent manner (5-15 μg/mL) (p < 0.05), while it exhibited no toxic effect on normal chick embryo fibroblasts. The antiproliferative mechanism of LFP(91-3)A2 was found to be associated with inducing cell apoptosis by flow cytometry analysis and transmission electron microscopy. The LFP(91-3)A2 is a novel protein from Lentinula edodes with tumor-suppressive activity via inducing apoptosis of tumor cells without toxicity on normal cells and may be beneficial to natural products in clinical treatment.
我们从香菇 C（91-3）液体菌丝体培养上清液中纯化并分离出一种具有抗肿瘤作用的新型蛋白（LFP（91-3）A2）。LFP（91-3）A2 通过（NH4）2SO4 沉淀、离子交换层析（DEAE-纤维素）和凝胶过滤层析（Sephacryl S-200HR）进行纯化。SDS-PAGE 和 MALDI-TOF/MS 分析 Mascot 搜索显示 LFP（91-3）A2 是一种新的蛋白质，表观分子量为 26 kDa。体外 MTT 测定法评估了对肿瘤细胞增殖的影响，LFP（91-3）A2 以剂量依赖性方式（5-15 μg/mL）明显降低肿瘤细胞生长（p < 0.05），而对正常鸡胚成纤维细胞无毒性作用。通过流式细胞术分析和透射电子显微镜发现，LFP（91-3）A2 的抗增殖机制与诱导细胞凋亡有关。LFP（91-3）A2 是一种来自香菇的新型蛋白，通过诱导肿瘤细胞凋亡而具有肿瘤抑制活性，对正常细胞无毒性，可能有益于临床治疗中的天然产物。
