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Cordyceps pruinosa extracts induce apoptosis of HeLa cells by a caspase dependent pathway
Authors:Ho Gyoung Kim  Heesang Song  Deok Hyo Yoon  Byeong-Wook Song  Sang Min Park  Gi Ho Sung  Jae-Youl Cho  Hae Il Park  Sunga Choi  Won O Song  Ki-Chul Hwang  Tae Woong Kim
Affiliation:1. Department of Biochemistry, Kangwon National University, Chuncheon 200-701, Republic of Korea;2. Research Institute of Science for Aging, Yonsei University, Seoul 120-752, Republic of Korea;3. Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul 120-752, Republic of Korea;4. Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR 97331, USA;5. School of Bioscience & Biotechnology, Kangwon National University, Chuncheon 200-701, Republic of Korea;6. Division of Pharmacology, Kangwon National University, Chuncheon 200-701, Republic of Korea;g Department of Life Science, Hallym University, Chuncheon 200-702, Republic of Korea;h Department of Food Science and Human Nutrition, Michigan State University, East Lansing, MI 48824, USA;i Cardiovascular Research Institute, Yonsei University College of Medicine, Seoul 120-752, Republic of Korea
Abstract:

Aim of the study

Cordyceps is a parasitic fungus and has long been used as a traditional Chinese medicine to treat illnesses, promote longevity, increase athletic power, and relieve exhaustion and cancer. In this study, we reveal the mechanisms underlying apoptosis induced by Cordyceps pruinosa butanol fraction (CPBF) in the human cervical adenocarcinoma cell line, HeLa.

Materials and methods

Proliferation and apoptosis of cells were examined by MTT assay, DNA fragmentation, phosphatidyl serine distribution assay, Western blot analysis, and immunocytochemistry. To determine the association between CPBF related apoptosis and ROS, electron spin resonance (ESR) trapping experiments were used.

Results

CPBF inhibited proliferation and induced apoptosis in HeLa cells in a dose-dependent manner using a MTT assay, DNA fragmentation, and a phosphatidyl serine distribution assay. Western blot analysis showed that apoptosis in HeLa cells was caspase-3- and -9-dependent. Proteolytic cleavage of PARP and the release of cytochrome c from the mitochondria into the cytosol were significantly increased and the Bcl-2/Bax protein ratio was decreased. Apoptosis induced by CPBF was not prevented by various antioxidants.

Conclusions

These results indicate that apoptotic effects of CPBF on HeLa cells are mediated by mitochondria-dependent death-signaling pathway independent of reactive oxygen species, suggesting that CPBF might be effective as an anti-proliferative agent for cancer.
Keywords:CPBF, Cordyceps pruinosa butanol fraction   TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling   PARP, poly(ADP-ribose) polymerase   ROS, reactive oxygen species   ESR, electron spin resonance
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