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

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

doi:10.3831/kpi.2022.25.4.390

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

Journal of pharmacopuncture, v.25 no.4, 2022년, pp.390 - 395

Yu-Na Hwang (Department of Biological Sciences, College of Natural Sciences, Kangwon National University) , In-Seo Kwon (Department of Biological Sciences, College of Natural Sciences, Kangwon National University) , Han-Heom Na (Department of Biological Sciences, College of Natural Sciences, Kangwon National University) , Jin-Sung Park (Korean Pharmacopuncture Institute) , Keun-Cheol Kim (Department of Biological Sciences, College of Natural Sciences, Kangwon National University)

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Objectives: Sweet bee venom (sBV) is purified from Apis mellifera, 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 ㎍/mL of sBV. The MTT assay indicated that sBV treatment exhibited a growth inhibitory effect at a concentration over 5 ㎍/mL. On fluorescence activated cell sorting analysis, G0 dead cells were observed after G1 arrest at treatment concentrations up to 10 ㎍/mL. However, rapid cell rupture was observed at a concentration of 20 ㎍/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.

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그림 Figure 1. Morphological changes in sBV-treated A549 cells. A549 cells were treated with sBV at 5 μg/mL, 10 μg/mL, and 20 μg/mL concentrations for 24 hr. Morphological changes were observed with a phase contrast microscope. Acute cell rupture was observed with 20 μg sBV treatment.
그림 Figure 2. Growth inhibitory effect and FACS analysis in sBV-treated A549 cells. (A) A549 cells were treated with 5 μg/mL, 10 μg/mL, and 20 μg/mL of sBV for 72 hr. MTT assay was performed to examine cell proliferation. (B) FACS analysis was performed after sBV treatment at various concentrations. Cell death was observed after G1 arrest at a pharmacological concentration, but acute cell death was observed with 20 μg sBV treatment.
그림 Figure 3. Expression of cell death-related proteins in sBV-treated A549 cells. Western blot analysis was performed after sBV treatment in A549 cells. Cleaved forms of PARP-1 and Caspase 9 were increased, and p53 or Bax was also upregulated with sBV treatment in a dose-dependent manner.

참고문헌 (24)

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

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