Ribosome-inactivating Protein MAP30 Isolated from Momordica Charantia L. Induces Apoptosis in Hepatocellular Carcinoma Cells.

IF 2.5 4区 医学 Q3 ONCOLOGY
Yiping Zhou, Di Yang, Zihao Qiang, Yanfa Meng, Ruigang Li, Xiang Fan, Wei Zhao, Yao Meng
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Abstract

Background: Ribosome-inactivating proteins (RIPs) have been reported to exert antitumor and anti-virus activities. A recent patent CN202011568116.7 has developed a new method to prepare Momordica anti-HIV protein of 30 kDa (MAP30). MAP30 is a type I RIP, which kills various tumor cells through the N-glycosidase activity and irreversibly inhibits protein synthesis.

Objective: To assess the potential role of MAP30 in inducing apoptosis of human hepatocellular carcinoma HCC-LM3 cells and elucidate the molecular mechanism of MAP30.

Methods: CCK-8 assay was used to assess the proliferation of HCC-LM3 cells. Flow cytometry was used to measure the cycle, the level of ROS and apoptosis in HCC-LM3 cells. Western blots was used to measure protein levels.

Results: Treatment with MAP30 reduced survival and proliferation of human liver cancer HCCLM3 cells in a dose-dependent manner. PI staining showed cell cycle arrest in G0/G1 phase. Furthermore, MAP30 increased the level of ROS in HCC-LM3 cells in 24 h treatment. To further confirm the role of MAP30 in inducing cell apoptosis, immunoblotting was carried out to detect the change of apoptosis-related proteins including PARP poly (ADP-ribose) polymerase (PARP- 1), Casepase3 and Cleaved-Caspase9. We found that PARP-1 and Caspase-3 were downregulated, whereas Cleaved-Caspase9 was up-regulated in HCC-LM3 cells treated with MAP30.

Conclusion: This study indicated that MAP30 has the potential to be a novel therapeutic agent for human hepatocellular carcinoma.

从 Momordica Charantia L. 中分离出的核糖体失活蛋白 MAP30 能诱导肝细胞癌细胞凋亡。
背景:据报道,核糖体失活蛋白(RIPs)具有抗肿瘤和抗病毒活性。最近的一项专利 CN202011568116.7 开发了一种制备 30 kDa Momordica 抗艾滋病毒蛋白(MAP30)的新方法。MAP30 是一种 I 型 RIP,通过 N-糖苷酶活性杀死各种肿瘤细胞,并不可逆地抑制蛋白质合成:评估 MAP30 在诱导人肝癌 HCC-LM3 细胞凋亡中的潜在作用,并阐明 MAP30 的分子机制:采用 CCK-8 检测法评估 HCC-LM3 细胞的增殖情况。流式细胞术用于测量 HCC-LM3 细胞的周期、ROS 水平和凋亡。用 Western 印迹法测定蛋白质水平:结果:用 MAP30 处理人肝癌 HCCLM3 细胞可降低其存活率和增殖率,且呈剂量依赖性。PI 染色显示细胞周期停滞在 G0/G1 期。此外,MAP30 还能提高 HCC-LM3 细胞在 24 小时处理过程中的 ROS 水平。为了进一步证实 MAP30 在诱导细胞凋亡中的作用,我们采用免疫印迹法检测了 PARP 多(ADP-核糖)聚合酶(PARP-1)、Casepase3 和裂解-Caspase9 等细胞凋亡相关蛋白的变化。我们发现,在用MAP30处理的HCC-LM3细胞中,PARP-1和Caspase-3下调,而Cleaved-Caspase9上调:结论:这项研究表明,MAP30 有可能成为治疗人类肝细胞癌的新型药物。
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来源期刊
CiteScore
4.50
自引率
7.10%
发文量
55
审稿时长
3 months
期刊介绍: Aims & Scope Recent Patents on Anti-Cancer Drug Discovery publishes review and research articles that reflect or deal with studies in relation to a patent, application of reported patents in a study, discussion of comparison of results regarding application of a given patent, etc., and also guest edited thematic issues on recent patents in the field of anti-cancer drug discovery e.g. on novel bioactive compounds, analogs, targets & predictive biomarkers & drug efficacy biomarkers. The journal also publishes book reviews of eBooks and books on anti-cancer drug discovery. A selection of important and recent patents on anti-cancer drug discovery is also included in the journal. The journal is essential reading for all researchers involved in anti-cancer drug design and discovery. The journal also covers recent research (where patents have been registered) in fast emerging therapeutic areas/targets & therapeutic agents related to anti-cancer drug discovery.
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