Cordyceps pruinosa extracts induce apoptosis of HeLa cells by a caspase dependent pathway.
OBJECTIVE
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.


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.
Department of Biochemistry, Kangwon National University, Chuncheon 200-701, Republic of Korea.
Ho Gyoung
HG
Kim
Heesang
H
Song
Deok Hyo
DH
Yoon
Byeong-Wook
BW
Song
Sang Min
SM
Park
Gi Ho
GH
Sung
Jae-Youl
JY
Cho
Hae Il
HI
Park
Sunga
S
Choi
Won O
WO
Song
Ki-Chul
KC
Hwang
Tae Woong
TW
Kim
D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism*; D020169:Caspases / Q000378:metabolism*; D032661:Cordyceps / Q000378:metabolism*; D003574:Cytochrome c Group / Q000378:metabolism; D045304:Cytochromes c / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D008928:Mitochondria / Q000378:metabolism; D010936:Plant Extracts / Q000494:pharmacology*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D003574:Cytochrome c Group; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D051028:bcl-2-Associated X Protein; C076615:cytochrome c''; D045304:Cytochromes c; D011065:Poly(ADP-ribose) Polymerases; D053148:Caspase 3; D020169:Caspases