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甜蜂毒以剂量依赖性方式处理A549人肺癌细胞诱导双重细胞毒性反应。

Dual Cytotoxic Responses Induced by Treatment of A549 Human Lung Cancer Cells with Sweet Bee Venom in a Dose-Dependent Manner.

作者信息

Hwang Yu-Na, Kwon In-Seo, Na Han-Heom, Park Jin-Sung, Kim Keun-Cheol

机构信息

Department of Biological Sciences, College of Natural Sciences, Kangwon National University, Chuncheon, Republic of Korea.

Kangwon Center for System Imaging, Kangwon National University, Chuncheon, Republic of Korea.

出版信息

J Pharmacopuncture. 2022 Dec 31;25(4):390-395. doi: 10.3831/KPI.2022.25.4.390.

Abstract

OBJECTIVES

Sweet bee venom (sBV) is purified from , containing a high level of melittin-its main component. It has been used as a therapeutic agent for pain relief and anti-inflammation, as well as for treating neuronal abnormalities. Recently, there have been studies on the therapeutic application of sBV for anticancer treatment. In the present study, we investigated the pharmacological effect of sBV treatment in A549 human lung cancer cells.

METHODS

We used microscopic analysis to observe the morphological changes in A549 cells after sBV treatment. The MTT assay was used to examine the cytotoxic effect after dose-dependent sBV treatment. Molecular changes in sBV were evaluated by the expression of apoptosis marker proteins using western blot analysis.

RESULTS

Microscopic analysis suggested that the growth inhibitory effect occurred in a dose-dependent manner; however, cell lysis occurred at a concentration over 20 μg/mL of sBV. The MTT assay indicated that sBV treatment exhibited a growth inhibitory effect at a concentration over 5 μg/mL. On fluorescence activated cell sorting analysis, G0 dead cells were observed after G1 arrest at treatment concentrations up to 10 μg/mL. However, rapid cell rupture was observed at a concentration of 20 μg/mL. Western blot analysis demonstrated that sBV treatment modulated the expression of multiple cell death-related proteins, including cleaved-PARP, cleaved-caspase 9, p53, Bcl2, and Bax.

CONCLUSION

sBV induced cell death in A549 human lung cancer cells at a pharmacological concentration, albeit causing hemolytic cell death at a high concentration.

摘要

目的

甜蜂毒(sBV)是从[具体来源未提及]中纯化得到的,含有高水平的蜂毒肽——其主要成分。它已被用作缓解疼痛和抗炎的治疗剂,以及用于治疗神经元异常。最近,有关于sBV在抗癌治疗中的治疗应用研究。在本研究中,我们研究了sBV处理对A549人肺癌细胞的药理作用。

方法

我们使用显微镜分析观察sBV处理后A549细胞的形态变化。MTT法用于检测剂量依赖性sBV处理后的细胞毒性作用。通过蛋白质免疫印迹分析凋亡标记蛋白的表达来评估sBV的分子变化。

结果

显微镜分析表明生长抑制作用呈剂量依赖性;然而,当sBV浓度超过20μg/mL时会发生细胞裂解。MTT法表明sBV处理在浓度超过5μg/mL时表现出生长抑制作用。在荧光激活细胞分选分析中,在处理浓度高达10μg/mL时,G1期阻滞之后观察到G0期死亡细胞。然而,在浓度为20μg/mL时观察到细胞迅速破裂。蛋白质免疫印迹分析表明sBV处理调节了多种细胞死亡相关蛋白的表达,包括裂解的PARP、裂解的半胱天冬酶-9、p53、Bcl2和Bax。

结论

sBV在药理浓度下诱导A549人肺癌细胞死亡,尽管在高浓度下会导致溶血性细胞死亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0e55/9806155/fd6d70fecd61/jop-25-4-390-f1.jpg

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