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KMID : 0615720030200020098
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Jouranl of Korean Acupuncture and Moxibustion society
2003 Volume.20 No. 2 p.98 ~ p.111
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The Effects of Bee Venom & Melittin on Cell Death in Neuroblastoma Cell Line
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Abstract
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Objective : This study was designed to analyze the effects of bee venom and melittin on cell death I neuroblastoma cell line.
Methods : MTT assay, morphologic method, DNA fragmentation, flow cytometry, immunocytochemistry analysis. RT-PCR and Western blot were performed.
Results : The obtained results are summarized as follows:
1. The MTT assay demonstrated that neuroblastoma cell viability was significantly inhibitted dose-dependently by treatment with bee venom and melittin in comparison with control.
2 Cell culture demonstrated that control group proliferated highestly at he 5th day in comparison with the 4th day in bee venom and melittin group. and in bee venom and melitti group cell proliferation decreased 2.5 times than control group.
3. The morphologic study demonstrated that neuroblastoma cell showed apoptosis after treatment with bee venom and melittin for 6 hours using microscope.
4. The Flow cytometry demonstrated that apoptosis of neuroblastoma cell treated with bee venom and melittin was related with stop of cell cycle in stage of G_10/G_1.
5.DNA fragmenation demonstrated that neuroblastoma cell treated with bee venom and melittin showed DNA ladder below 1 Kbp.
6. Immnocytochemistry assay demonstrated that Fos and MAPK which rae related with cancer were down-regulated by treatment with bee venom and melittin.
7. RT-PCR analysis demonstrated that Fos and MAPK mRNA were transcripted. Fos was down-regulated form treatment with 5ug/ml bee venom and MAPK was down-regulated form 1ug/ml bee venom.
8. Western blot demonstrated that Fos was down-regulated from 1ug/ml bee venom whereas MAPK was expressed by 1 ug/ml bee venom but down-regulated by 10 ug/ml bee venom.
Conclusions : We found that some cancer related genes ware down-regulated by treatment with bee venom and melittin. Further study is needed for investigating the anti-cancer effect of bee venom and melittin.
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