PMID- 24659394
OWN - NLM
STAT- MEDLINE
DCOM- 20140626
LR  - 20211203
IS  - 1432-1335 (Electronic)
IS  - 0171-5216 (Linking)
VI  - 140
IP  - 6
DP  - 2014 Jun
TI  - Combination of Cl‑IB‑MECA with paclitaxel is a highly effective cytotoxic therapy 
      causing mTOR‑dependent autophagy and mitotic catastrophe on human melanoma cells.
PG  - 921-35
AB  - PURPOSE: Metastatic melanoma is the deadliest form of skin cancer. It is highly 
      resistant to conventional therapies,particularly to drugs that cause apoptosis as 
      the main anticancer mechanism. Recently, induction of autophagic cell death is 
      emerging as a novel therapeutic target for apoptotic-resistant cancers. We aimed 
      to investigate the underlying mechanisms elicited by the cytotoxic combination of 
      2-chloro-N(6)-(3-iodobenzyl)-adenosine-5'-N-methyluronamide(Cl-IB-MECA, a 
      selective A(3) adenosine receptor agonist; 10 muM) and paclitaxel (10 ng/mL) on 
      human C32 and A375 melanoma cell lines. METHODS: Cytotoxicity was evaluated using 
      3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide reduction, neutral 
      red uptake, and lactate dehydrogenase leakage assays, after 48-h incubation. 
      Autophagosome and autolysosome formation was detected by fluorescence through 
      monodansylcadaverine-staining and CellLight((R)) Lysosomes-RFP-labelling, 
      respectively. Cell nuclei were visualized by Hoechst staining, while levels of 
      p62 were determined by an ELISA kit. Levels of mammalian target of rapamycin 
      (mTOR) and the alterations of microtubule networks were evaluated by 
      immunofluorescence. RESULTS: We demonstrated, for the first time, that the 
      combination of Cl-IB-MECA with paclitaxel significantly increases cytotoxicity, 
      with apoptosis and autophagy the major mechanisms involved in cell death. 
      Induction of autophagy, using clinically relevant doses,was confirmed by 
      visualization of autophagosome and autolysosome formation, and downregulation of 
      mTOR and p62 levels. Caspase-dependent and caspase-independent mitotic 
      catastrophe evidencing micro- and multinucleation was also observed in cells 
      exposed to our combination. CONCLUSIONS: The combination of Cl-IB-MECA and 
      paclitaxel causes significant cytotoxicity on two melanoma cell lines through 
      multiple mechanisms of cell death. This multifactorial hit makes this therapy 
      very promising as it will help to avoid melanoma multiresistance to chemotherapy 
      and therefore potentially improve its treatment.
FAU - Soares, Ana S
AU  - Soares AS
FAU - Costa, Vera M
AU  - Costa VM
FAU - Diniz, Carmen
AU  - Diniz C
FAU - Fresco, Paula
AU  - Fresco P
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Germany
TA  - J Cancer Res Clin Oncol
JT  - Journal of cancer research and clinical oncology
JID - 7902060
RN  - 0 (Adenosine A3 Receptor Agonists)
RN  - 0 (Antineoplastic Agents, Phytogenic)
RN  - EC 2.7.1.1 (MTOR protein, human)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
RN  - K72T3FS567 (Adenosine)
RN  - P88XT4IS4D (Paclitaxel)
RN  - Z07JR07J6C (2-chloro-N(6)-(3-iodobenzyl)adenosine-5'-N-methyluronamide)
SB  - IM
MH  - Adenosine/*analogs & derivatives/pharmacology
MH  - Adenosine A3 Receptor Agonists/pharmacology
MH  - Antineoplastic Agents, Phytogenic/pharmacology
MH  - Apoptosis/drug effects
MH  - Autophagy/*drug effects
MH  - Blotting, Western
MH  - Cell Proliferation/drug effects
MH  - *Drug Synergism
MH  - Drug Therapy, Combination
MH  - Fluorescent Antibody Technique
MH  - Humans
MH  - Melanoma/drug therapy/metabolism/*pathology
MH  - Mitosis/*drug effects
MH  - Paclitaxel/*pharmacology
MH  - TOR Serine-Threonine Kinases/*metabolism
MH  - Tumor Cells, Cultured
EDAT- 2014/03/25 06:00
MHDA- 2014/06/27 06:00
CRDT- 2014/03/25 06:00
PHST- 2014/02/26 00:00 [received]
PHST- 2014/03/07 00:00 [accepted]
PHST- 2014/03/25 06:00 [entrez]
PHST- 2014/03/25 06:00 [pubmed]
PHST- 2014/06/27 06:00 [medline]
AID - 10.1007/s00432-014-1645-z [doi]
PST - ppublish
SO  - J Cancer Res Clin Oncol. 2014 Jun;140(6):921-35. doi: 10.1007/s00432-014-1645-z.