Crude extracts of Euchresta formosana radix induce cytotoxicity and apoptosis in human hepatocellular carcinoma cell line (Hep3B).
In this study, the effects of 95% ethanol extracts of Euchresta formosana radix (EFR) on the cell cycle and apoptosis in human hepatocellular carcinoma (HCC) Hep3B cells were investigated. The results indicated that EFR decreased DNA synthesis and viable Hep3B cell numbers in a concentration-dependent manner. EFR induced a p21- and p27-dependent cell cycle arrest in S-phase and apoptosis of the Hep3B cells. The induction of apoptosis by EFR treatment was also confirmed by DAPI staining. EFR inhibited cyclin-dependent kinase (CDK)-1 and -2 expression and decreased cyclin B1 and E levels, resulting in S-phase arrest. EFR induced reactive oxygen species (ROS) production followed by endoplasmic reticulum (ER) stress that was based on the increase of GADD153 and GRP78 which led to the release of Ca2+ in the Hep3B cells. The EFR-promoted apoptosis was associated with increasing activation of caspases 3, 7, and 9 and enhanced poly(ADP-ribose) polymerase cleavage and increased expression of p21(CIP1/WAF1), p27(KIP1), Bax and Bad. Furthermore, the levels of Bcl-xl decreased after EFR treatment. Alteration of these key anti- and pro-apoptotic proteins could contribute to the increase in p53-independent apoptosis that was observed in the Hep3B cells.
Hsu
Shu-Chun
SC
Graduate Institute of Chinese Pharmaceutical Sciences, China Medical University, Taichung 404, Taiwan, ROC.
Kuo
Chao-Lin
CL
Lin
Jing-Pin
JP
Lee
Jau-Hong
JH
Lin
Chin-Chung
CC
Su
Chin-Cheng
CC
Lin
Hui-Ju
HJ
Chung
Jing-Gung
JG
D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D053148:Caspase 3 / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D003593:Cytoplasm / Q000378:metabolism; D000091342:Endoplasmic Reticulum Chaperone BiP; D000431:Ethanol / Q000737:chemistry; D007887:Fabaceae / Q000737:chemistry*; D005456:Fluorescent Dyes; D006801:Humans; D007211:Indoles; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D010936:Plant Extracts / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism
D000091342:Endoplasmic Reticulum Chaperone BiP; D005456:Fluorescent Dyes; C000717247:HSPA5 protein, human; D007211:Indoles; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D000431:Ethanol; C007293:DAPI; D053148:Caspase 3; D002118:Calcium