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从苦瓜中提取的两种核糖体失活蛋白α-苦瓜毒蛋白(α-MMC)和MAP30,可诱导人肺癌A549细胞的细胞周期阻滞和凋亡。

Α-MMC and MAP30, two ribosome-inactivating proteins extracted from Momordica charantia, induce cell cycle arrest and apoptosis in A549 human lung carcinoma cells.

作者信息

Fan Xiang, He Lingli, Meng Yao, Li Gangrui, Li Linli, Meng Yanfa

机构信息

Key Laboratory of China Ministry Education for Bio‑Resources and Eco‑Environment/Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, Sichuan 610065, P.R. China.

School of Medical Laboratory Science, Chengdu Medical College, Chengdu, Sichuan 610500, P.R. China.

出版信息

Mol Med Rep. 2015 May;11(5):3553-8. doi: 10.3892/mmr.2015.3176. Epub 2015 Jan 9.

Abstract

α‑Momorcharin (α‑MMC) and momordica anti‑human immunodeficiency virus protein (MAP30), produced by Momordica charantia, are ribosome‑inactivating proteins, which have been reported to exert inhibitory effects on cultured tumor cells. In order to further elucidate the functions of these agents, the present study aimed to investigate the effects of α‑MMC and MAP30 on cell viability, the induction of apoptosis, cell cycle arrest, DNA integrity and superoxide dismutase (SOD) activity. α‑MMC and MAP30 were purified from bitter melon seeds using ammonium sulfate precipitation in combination with sulfopropyl (SP)‑sepharose fast flow, sephacryl S‑100 and macro‑Cap‑SP chromatography. MTT, flow cytometric and DNA fragmentation analyses were then used to determine the effects of α‑MMC and MAP30 on human lung adenocarcinoma epithelial A549 cells. The results revealed that A549 cells were sensitive to α‑MMC and MAP30 cytotoxicity assays in vitro. Cell proliferation was significantly suppressed following α‑MMC and MAP30 treatment in a dose‑ and time‑dependent manner; in addition, the results indicated that MAP30 had a more potent anti‑tumor activity compared with that of α‑MMC. Cell cycle arrest in S phase and a significantly increased apoptotic rate were observed following treatment with α‑MMC and MAP30. Furthermore, DNA integrity analysis revealed that the DNA of A549 cells was degraded following treatment with α‑MMC and MAP30 for 48 h. The pyrogallol autoxidation method and nitrotetrazolium blue chloride staining were used to determine SOD activity, the results of which indicated that α‑MMC and MAP30 did not possess SOD activity. In conclusion, the results of the present study indicated that α‑MMC and MAP30 may have potential as novel therapeutic agents for the prophylaxis and treatment of cancer.

摘要

α-苦瓜素(α-MMC)和苦瓜抗人免疫缺陷病毒蛋白(MAP30)由苦瓜产生,是核糖体失活蛋白,据报道对培养的肿瘤细胞具有抑制作用。为了进一步阐明这些药物的功能,本研究旨在探讨α-MMC和MAP30对细胞活力、凋亡诱导、细胞周期阻滞、DNA完整性和超氧化物歧化酶(SOD)活性的影响。采用硫酸铵沉淀结合磺丙基(SP)-琼脂糖快速流动、Sephacryl S-100和大体积Cap-SP色谱法从苦瓜种子中纯化α-MMC和MAP30。然后使用MTT、流式细胞术和DNA片段分析来确定α-MMC和MAP30对人肺腺癌上皮A549细胞的影响。结果显示,A549细胞在体外对α-MMC和MAP30细胞毒性试验敏感。α-MMC和MAP30处理后,细胞增殖以剂量和时间依赖性方式显著受到抑制;此外,结果表明MAP30比α-MMC具有更强的抗肿瘤活性。用α-MMC和MAP30处理后,观察到细胞周期阻滞在S期,凋亡率显著增加。此外,DNA完整性分析显示,用α-MMC和MAP30处理48小时后,A549细胞的DNA被降解。采用邻苯三酚自氧化法和氯化硝基四氮唑蓝染色法测定SOD活性,结果表明α-MMC和MAP30不具有SOD活性。总之,本研究结果表明,α-MMC和MAP30可能具有作为癌症预防和治疗新型治疗药物的潜力。

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