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ICD - 85(毒液衍生肽)对人乳腺腺癌和正常人皮肤成纤维细胞的细胞毒性及抗癌作用。

Cytotoxic and Anticancer Effects of ICD-85 (Venom Derived Peptides) in Human Breast Adenocarcinoma and Normal Human Dermal Fibroblasts.

作者信息

Kheirandish Zarandi Peyman, Zare Mirakabadi Abbas, Sotoodehnejadnematalahi Fattah

机构信息

Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.

Department of Venomous Animal and Antivenom Production, Razi Vaccine and Serum Research Institute, Agriculture Research Education and Extension Organization (AREEO), Karaj, Iran.

出版信息

Iran J Pharm Res. 2019 Winter;18(1):232-240.

Abstract

ICD-85 (venom derived peptides) has anti-proliferative effect and anti- angiogenesis activity on cancer cells. This study was performed to test the effect of ICD-85, on Human breast adenocarcinoma (MCF-7) and normal Human Dermal Fibroblasts (HDF) cell lines. In this experimental study, Mitochondrial activity, Neutral red uptake, Lactate dehydrogenase (cell necrosis), and cell morphology were assessed under unexposed and ICD-85 exposed conditions. Caspase-9 colorimetric assay kit was used to determine caspase protease activity. Morphological changes in MCF-7 cells on treatment with ICD-85 compared with untreated MCF-7 cells are consistent with characterizing the features of apoptosis such as granulation and cell rounding which finally results in the generation of apoptotic bodies. In contrast, this difference was not observed in normal cells. In MTT assay, ICD-85 induced dose dependent manner cytotoxic effects on MCF-7 cells which were confirmed by neutral red assay. The results showed that inhibitory concentration 50% (IC50) value of ICD-85 for MCF-7 cells at 24 h was 36.45 ± 0.38 μg/mL. However, when HDF cells were exposed to ICD-85, no significant elevation of LDH release were observed at concentrations below 20 μg/mL. The apoptosis-induction of ICD-85 on MCF-7 cell was found to be through activation of caspase-9 which was 13 fold greater than unexposed cell. This study showed that ICD-85 induced apoptosis in MCF-7 cell line through caspase activation and hence it can be considered for further investigation to use ICD-85 as a potential therapy for breast cancer.

摘要

ICD - 85(毒液衍生肽)对癌细胞具有抗增殖作用和抗血管生成活性。本研究旨在测试ICD - 85对人乳腺腺癌(MCF - 7)和正常人皮肤成纤维细胞(HDF)细胞系的影响。在本实验研究中,在未暴露和ICD - 85暴露条件下评估线粒体活性、中性红摄取、乳酸脱氢酶(细胞坏死)和细胞形态。使用半胱天冬酶 - 9比色测定试剂盒来确定半胱天冬酶蛋白酶活性。与未处理的MCF - 7细胞相比,用ICD - 85处理的MCF - 7细胞的形态变化与凋亡特征(如颗粒化和细胞变圆)一致,最终导致凋亡小体的产生。相比之下,在正常细胞中未观察到这种差异。在MTT试验中,ICD - 85对MCF - 7细胞诱导了剂量依赖性的细胞毒性作用,这通过中性红试验得到证实。结果表明,ICD - 85在24小时时对MCF - 7细胞的半数抑制浓度(IC50)值为36.45±0.38μg/mL。然而,当HDF细胞暴露于ICD - 85时,在浓度低于20μg/mL时未观察到乳酸脱氢酶释放的显著升高。发现ICD - 85对MCF - 7细胞的凋亡诱导是通过半胱天冬酶 - 9的激活,其比未暴露细胞高13倍。本研究表明,ICD - 85通过半胱天冬酶激活在MCF - 7细胞系中诱导凋亡,因此可以考虑进一步研究将ICD - 85用作乳腺癌的潜在治疗方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e2fb/6487428/4e1fbdf7b734/ijpr-18-232-g001.jpg

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