臨床病理学ジャーナル

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Nuclear Area Factor as a Novel Estimate for Apoptosis in Oral Squamous Cell Carcinoma -Treated Cell Line: A Comparative in-vitro Study with DNA Fragmentation Assay

Nermeen S. Afifi, Ehab S. Abdel-Hamid, Houry M. Baghdadi and Ali F. Mohamed

Aims and objectives:The aims of the present study were to introduce a novel modality for estimation of the anti-neoplastic efficacy of chemotherapeutic agents and to compare this technique with one of the most widely used techniques, the DNA fragmentation assay.

Methods:Squamous cell carcinoma cell line HEp-2 was utilized in the present study. The cytotoxic drug used in this study was gossypol-acetic acid. Microscopic slides of each drug concentration at definite post-treatment durations were photomicrographed and analyzed using image anazlysis software for the estimation of nuclear area factor. Data were then statistically analyzed. Internucleosomal DNA fragmentation was investigated to assess whether the morphological changes induced by the drug depended on activation of the apoptotic pathway.

Results:The data recorded revealed a decrease in the mean values of NAF of HEp-2 cells treated with different concentrations of gossypol over different durations when compared to control cells. DNA fragmentation was not observed in the control groups over different durations of incubation. The study of internucleosomal DNA fragmentation in HEp-2 cells treated with different concentrations of gossypol over different durations revealed absence of DNA laddering that indicated DNA fragmentation with either 50μM or 100μM gossypol concentrations after until 24 hours of incubation with 50μM gossypol concentration.

Conclusions:These results suggest that estimation of nuclear area factor (NAF) is a sensitive predictor of the early apoptotic effect of anti-cancer therapy. DNA fragmentation assay is a useful tool for detection of the late apoptotic changes that discriminate apoptotic effects of anti-cancer drugs from necrotic ones.