一种85个氨基酸的多肽,与热休克因子结合蛋白1同源,体外对MG-63骨肉瘤细胞具有抗增殖活性。

IF 0.4 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Asian Biomedicine Pub Date : 2022-08-01 DOI:10.2478/abm-2022-0024
Rui Ding, Ming He, Huoying Huang, Jing Chen, Mingxing Huang, Yonghui Su
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引用次数: 0

摘要

背景:有毒节肢动物的毒液中含有对肿瘤细胞具有抗增殖潜能的物质。目的:鉴定一种对肿瘤细胞具有抗增殖活性的多肽,并阐明其分子机制。方法:采用凝胶过滤和离子交换色谱法从蚁甲蛋白提取物中纯化出具有抗MG-63人骨肉瘤细胞增殖活性的多肽。对该多肽进行了测序,并在体外一系列条件下测试了其抗增殖活性的稳定性。采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑(MTT)法测定该多肽对MG-63骨肉瘤细胞和MC3T3-E1小鼠颅骨成骨细胞的抗增殖活性,并将其作为非肿瘤对照。我们采用western blotting方法比较MG-63骨肉瘤细胞及其小鼠同源物在MC3T3-E1成骨细胞中经antlion抗增殖多肽(ALAPP)处理后热休克转录因子1 (HSF1)、热休克蛋白90 (HSP90)、细胞周期蛋白依赖性激酶4 (CDK4)和蛋白激酶B α (ATK1)的表达水平。结果:85个氨基酸组成的ALAPP与人热休克因子结合蛋白1 (HSBP1)的序列同源性为56%。多肽的抗增殖活性对温度、pH和金属离子相对不敏感。与对MC3T3-E1成骨细胞的作用相比,ALAPP对MG-63骨肉瘤细胞具有较强的浓度依赖性抗增殖活性。ALAPP在MC3T3-EL成骨细胞中显著上调HSF1的表达,而在MG-63骨肉瘤中无显著上调。ALAPP显著下调MG-63骨肉瘤中HSP90、CDK4和AKT1的表达,而在成骨细胞中无明显下调作用。结论:ALAPP对MG-63骨肉瘤细胞具有显著的抗增殖活性,但对非肿瘤性MC3T3-E1成骨细胞没有明显的抗增殖活性。我们推测,非肿瘤细胞可能通过上调HSF1,使其HSP90、CDK4和AKT1的表达维持在一个相对恒定的水平,从而逃避ALAPP的抗增殖作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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An 85-amino-acid polypeptide from Myrmeleon bore larvae (antlions) homologous to heat shock factor binding protein 1 with antiproliferative activity against MG-63 osteosarcoma cells in vitro.

Background: Venomous arthropods have substances in their venom with antiproliferative potential for neoplastic cells.

Objectives: To identify a polypeptide from Myrmeleon bore (antlion) with antiproliferative activity against neoplastic cells, and to elucidate the molecular mechanism of the activity.

Methods: We used gel filtration and ion exchange chromatography to purify a polypeptide with antiproliferative activity against MG-63 human osteosarcoma cells from a proteinaceous extract of antlion. The polypeptide was sequenced and the stability of its antiproliferative activity was tested under a range of conditions in vitro. An 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the antiproliferative activity of the polypeptide against the MG-63 osteosarcoma cells and MC3T3-E1 mouse calvarial osteoblasts, which were used as a non-neoplastic control. We used western blotting to compare the levels of expression of heat shock transcription factor 1 (HSF1), heat shock protein 90 (HSP90), cyclin-dependent kinase 4 (CDK4), and protein kinase B alpha (ATK1) in MG-63 osteosarcoma cells and their mouse homologs in MC3T3-E1 osteoblasts after their treatment with the antlion antiproliferative polypeptide (ALAPP).

Results: The 85-amino-acid ALAPP has a 56% sequence identity with the human heat shock factor binding protein 1 (HSBP1). The antiproliferative activity of the polypeptide is relatively insensitive to temperature, pH, and metal ions. ALAPP has a strong concentration-dependent antiproliferative activity against MG-63 osteosarcoma cells compared with its effect on MC3T3-E1 osteoblasts. ALAPP significantly upregulates the expression of HSF1 in MC3T3-EL osteoblasts, but not in MG-63 osteosarcoma. ALAPP significantly downregulated the expression of HSP90, CDK4, and AKT1 expression in MG-63 osteosarcoma, but not in the osteoblasts.

Conclusions: ALAPP has significant antiproliferative activity against MG-63 osteosarcoma cells, but not nonneoplastic MC3T3-E1 osteoblasts. We speculate that non-neoplastic cells may evade the antiproliferative effect of ALAPP by upregulating HSF1 to maintain their HSP90, CDK4, and AKT1 expression at a relatively constant level.

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来源期刊
Asian Biomedicine
Asian Biomedicine 医学-医学:研究与实验
CiteScore
1.20
自引率
0.00%
发文量
24
审稿时长
6-12 weeks
期刊介绍: Asian Biomedicine: Research, Reviews and News (ISSN 1905-7415 print; 1875-855X online) is published in one volume (of 6 bimonthly issues) a year since 2007. [...]Asian Biomedicine is an international, general medical and biomedical journal that aims to publish original peer-reviewed contributions dealing with various topics in the biomedical and health sciences from basic experimental to clinical aspects. The work and authorship must be strongly affiliated with a country in Asia, or with specific importance and relevance to the Asian region. The Journal will publish reviews, original experimental studies, observational studies, technical and clinical (case) reports, practice guidelines, historical perspectives of Asian biomedicine, clinicopathological conferences, and commentaries Asian biomedicine is intended for a broad and international audience, primarily those in the health professions including researchers, physician practitioners, basic medical scientists, dentists, educators, administrators, those in the assistive professions, such as nurses, and the many types of allied health professionals in research and health care delivery systems including those in training.
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