Combined cytotoxic action of paclitaxel and ceramide against the human Tu138 head and neck squamous carcinoma cell line.
OBJECTIVE
Paclitaxel, a chemotherapeutic agent used in the treatment of recalcitrant ovarian and breast as well as other neoplasms, is being investigated for the treatment of squamous cell carcinoma of the head and neck. Our previous studies have demonstrated that exogenously added ceramide enhances apoptosis in paclitaxel-exposed human leukemic cells. In this study, we showed that exogenous ceramide augmented paclitaxel-induced apoptosis in Tu138 cells in vitro when added simultaneously in combination with the paclitaxel.


METHODS
The combined cytotoxic effects of paclitaxel and ceramide exposure against Tu138 cells were assessed by an MTT dye assay, cell cycle analysis, TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) assay, and isobologram analysis for synergistic activity.


RESULTS
The MTT dye assay results indicated augmentation of time- and concentration-dependent paclitaxel-mediated cell cytotoxicity by simultaneous ceramide treatment. Paclitaxel treatment of Tu138 cells also resulted in an accumulation of cells in the G2-M phase of the cell cycle. This paclitaxel-mediated G2-M phase accumulation decreased significantly with the addition of ceramide, indicating that combined paclitaxel/ceramide treatment resulted in the elimination of Tu138 cells from the S and/or G2-M phases of the cell cycle. Furthermore, ceramide enhancement of paclitaxel-mediated apoptosis was also detected by the TUNEL assay.


CONCLUSIONS
Our results suggest that paclitaxel/ceramide combination therapy may be an attractive alternative to conventional methods of chemotherapy for head and neck cancer, and should be further explored.
Mehta
S
S
Department of Surgery, Roger Williams Medical Center, Boston University, Providence, Rhode Island 02908, USA.
Blackinton
D
D
Omar
I
I
Kouttab
N
N
Myrick
D
D
Klostergaard
J
J
Wanebo
H
H
D002294:Carcinoma, Squamous Cell; D002453:Cell Cycle / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects*; D002518:Ceramides / Q000633:toxicity*; D004357:Drug Synergism; D016193:G1 Phase; D016195:G2 Phase; D006258:Head and Neck Neoplasms; D006801:Humans; D020287:In Situ Nick-End Labeling; D008938:Mitosis; D017239:Paclitaxel / Q000633:toxicity*; D016192:Resting Phase, Cell Cycle; D014407:Tumor Cells, Cultured
D002518:Ceramides; C101954:N-caproylsphingosine; D017239:Paclitaxel