Methylated actinomycin D, a novel actinomycin D analog induces apoptosis in HepG2 cells through Fas- and mitochondria-mediated pathways
Article first published online: 20 JUL 2012
© 2012 Wiley Periodicals, Inc.
Volume 52, Issue 12, pages 983–996, December 2013
How to Cite
Chen, Y., Liu, J., Yuan, B., Cao, C., Qin, S., Cao, X., Bian, G., Wang, Z. and Jiang, J. (2013), Methylated actinomycin D, a novel actinomycin D analog induces apoptosis in HepG2 cells through Fas- and mitochondria-mediated pathways. Mol. Carcinog., 52: 983–996. doi: 10.1002/mc.21943
Disclosure information: None of the authors have anything to declare.
- Issue published online: 19 NOV 2013
- Article first published online: 20 JUL 2012
- Manuscript Accepted: 26 JUN 2012
- Manuscript Revised: 11 MAY 2012
- Manuscript Received: 9 DEC 2011
- Program of Natural Science Foundation of the Jiangsu Higher Education Institutions of China. Grant Number: 11KJB310011
- Chinese National Nature Science Foundation. Grant Number: 31170605, 31000005
- Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) (Chemicobiology)
- Outstanding Scientific and Technological Innovation
- methylated actinomycin D;
- HepG2 cells
Actinomycin D (Act D), a well known of clinical antitumor drug, has been used for the treatment of some highly malignant tumors, however, the clinical application was limited by its extreme cytotoxicity. In the present study, we reported that methylated actinomycin D (mAct D), a novel actinomycin D analog isolated from Streptomyces sp. KLBMP 2541 in our previous study, could not only exert stronger inhibitory effects on several human cancer cells than Act D in dose- and time-dependent manner at ng concentrations, especially on HepG2 cells, but also lower cytotoxicity in normal cells (HL-7702). Base on these results, HepG2 cells were treated for further study to illustrate the potential mechanism of mAct D. The results of nuclei morphology examination, DNA fragmentation detection, sub-G1 analysis, annexin V-FITC/PI staining and activation of caspase-3 indicated mAct D significantly induced HepG2 cells apoptosis. Semiquantitative RT-PCR and Western blot analysis revealed that mAct D induced apoptosis in HepG2 cells through mitochondria-dependent pathway by increasing levels of caspase-9, Bax, Bak while decreasing levels of Bcl-2, Bid, and Fas-dependent pathway by increasing levels of Fas, FasL, FADD, and caspase-8. Subsequently, pretreatment with specific inhibitor of caspase-8 Z-LEHD-FMK and caspase-9 Z-LEHD-FMK significantly attenuated caspase-3 activity, the cleavage of caspase-3 and PARP, meanwhile increased the cell viability. In addition, p53 and mitochondrial transcription factor A (mtTFA) were also upregulated. Taken together, ng concentrations mAct D induces the apoptosis of HepG2 through Fas- and mitochondria-mediated pathway and presents a potential novel alternative agent for the treatment of human hepatic carcinoma. © 2012 Wiley Periodicals, Inc.