Emission spectral analysis of caspase-3 activation during artesunate (ART)-induced apoptosis of human lung adenocarcinoma cell

Artesunate (ART), a semi-synthetic derivative of the sesquiterpene artemisinin extracted from the Chinese herb Artemisia annua, exerts a broad spectrum of clinical activity against human cancers. Artemisinin-derivative combination chemotherapy is recommended by WHO since it acts rapidly and is well tolerated and particularly effective. In present investigation, we used CKK-8 assay to assess the inhibitory effects of ART on human lung adenocarcinoma (ASTC-a-1) cells. Apoptotic activity of ART in ASTC-a-1 cells was detected by means of nuclear staining with Hoechst33258. In order to monitor the activity of caspase-3 during ART-induced ASTC-a-1 cells apoptosis, the dynamical emission spectra of SCAT3, a FRET plasmid based on GFPs, were performed inside living cell expressed stably with SCAT3 after ART treatment. The results showed that (1) ART could inhibit ASTC-a-1 cells proliferation in a dose-dependent manner; (2) chromatin condensation was observed after ART treatment for 48 h; (3) the SCAT3 inside living cells were cleaved after ART treatment for 48 h, implying that caspase-3 was involved in the ART-induced apoptosis.

[1]  M MacFarlane,et al.  Distinct Caspase Cascades Are Initiated in Receptor-mediated and Chemical-induced Apoptosis* , 1999, The Journal of Biological Chemistry.

[2]  D. Green.  Apoptotic Pathways The Roads to Ruin , 1998, Cell.

[3]  Da Xing,et al.  Bid Is Required in NPe6‐PDT‐induced Apoptosis , 2007, Photochemistry and photobiology.

[4]  T. Efferth,et al.  The anti-malarial artesunate is also active against cancer. , 2001, International journal of oncology.

[5]  P. Renard,et al.  Synthesis and cytotoxicity of artemisinin derivatives containing cyanoarylmethyl group. , 2001, European journal of medicinal chemistry.

[6]  Da Xing,et al.  Low‐power laser irradiation activates Src tyrosine kinase through reactive oxygen species‐mediated signaling pathway , 2008, Journal of cellular physiology.

[7]  Da Xing,et al.  Bid is not required for Bax translocation during UV-induced apoptosis. , 2007, Cellular signalling.

[8]  Da Xing,et al.  Single Cell FRET Imaging for Determination of Pathway of Tumor Cell Apoptosis Induced by Photofrin-PDT , 2006, Cell cycle.

[9]  Shaoqun Zeng,et al.  TRAIL-induced apoptosis proceeding from caspase-3-dependent and -independent pathways in distinct HeLa cells. , 2006, Biochemical and biophysical research communications.

[10]  Thomas Efferth,et al.  mRNA expression profiles for the response of human tumor cell lines to the antimalarial drugs artesunate, arteether, and artemether. , 2002, Biochemical pharmacology.

[11]  Takeharu Nagai,et al.  Spatio-temporal activation of caspase revealed by indicator that is insensitive to environmental effects , 2003, The Journal of cell biology.

[12]  C. Winterford,et al.  Apoptosis. Its significance in cancer and cancer Therapy , 1994, Cancer.

[13]  Jianhua Xu,et al.  Quantitative detection of the ligand-dependent interaction between the androgen receptor and the co-activator, Tif2, in live cells using two color, two photon fluorescence cross-correlation spectroscopy , 2007, European Biophysics Journal.

[14]  Da Xing,et al.  Detection of caspase-3 activation in single cells by fluorescence resonance energy transfer during photodynamic therapy induced apoptosis. , 2006, Cancer letters.