L-ascorbic acid induces apoptosis in human laryngeal epidermoid Hep-2 cells by modulating the nuclear factor kappa-light-chain-enhancer of activated B cells/ mitogen-activated protein kinase/Akt signa

서지정보

ㆍ발행기관 : 대한구강생물학회 ㆍ수록지정보 : International Journal of Oral Biology / 45권 / 4호
ㆍ저자명 : Jung-Sun Park, Yoon-Jung Kim, Sam Young Park, Kyung-Yi Chung, Sang-Jin Oh, Won-Jae Kim, Ji-Yeon Jung

목차

Introduction
Materials and Methods
1. Materials
2. Cell culture and cell viability assay
3. 4′,6-diamidino-2-phenylindole (DAPI) staining
4. Flow cytometry
5. Caspase activity and inhibition of apoptosis with acaspase inhibitor
6. Protein extraction and Western blot analysis
7. Extraction of nuclear protein
8. Electrophoretic mobility shift assay (EMSA)
9. Statistical analyses
Results
1. L-AA induced apoptotic cell death in humanlaryngeal Hep-2 cancer cells
2. L-AA induced the release of cytochrome c frommitochondria into the cytosol and upregulated AIFand EndoG levels
3. L-AA-induced apoptosis was caspase dependentin Hep-2 cells
4. L-AA-induced apoptosis is mediated via NF-κB,MAPK, and Akt signal pathways
Discussion
Acknowledgements
Conflicts of Interest
References

영어 초록

L-ascorbic acid (L-AA; vitamin C) induces apoptosis in cancer cells. This study aimed to elucidate the molecular mechanisms of L-AA-induced apoptosis in human laryngeal epidermoid carcinoma Hep-2 cells. L-AA suppressed the viability of Hep-2 cells and induced apoptosis, as shown by the cleavage and condensation of nuclear chromatin and increased number of Annexin V-positive cells. L-AA decreased Bcl-2 protein expression but upregulated Bax protein levels. In addition, cytochrome c release from the mitochondria into the cytosol and activation of caspase-9, -8, and -3 were enhanced by L-AA treatment. Furthermore, apoptosis-inducing factor (AIF) and endonuclease G (EndoG) were translocated into the nucleus during apoptosis of L-AA-treated Hep-2 cells. L-AA effectively inhibited the constitutive nuclear factor-κB (NF-κB) activation and attenuated the nuclear expression of the p65 subunit of NF-κB. Interestingly, L-AA treatment of Hep-2 cells markedly activated Akt and mitogen-activated protein kinase (MAPK; extracellular signal-regulated kinase 1/2, p38, and c-Jun N-terminal kinase [JNK]) and and LY294002 (Akt inhibitor), SB203580 (p38 inhibitor) or SP600125 (a JNK inhibitor) decreased the levels of Annexin V-positive cells. These results suggested that L-AA induces the apoptosis of Hep-2 cells via the nuclear translocation of AIF and EndoG by modulating the Bcl- 2 family and MAPK/Akt signaling pathways.

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