NUCLEAR AREA FACTOR VERSUS DNA FRAGMENTATION ASSAY FOR ESTIMATION OF APOPTOSIS IN SQUAMOUS CELL CARCINOMA -TREATED CELL LINE: A COMPARATIVE IN VITRO STUDY

Esraa Ali Mahmoud Bersy, Seham Ahmad Abdel Ghani And Houry Moustafa Baghdadi,

ABSTRACT
Aim And
Objectives: The Aim Of The Present Study Is To Introduce A Novel Modality For Estimation Of The Anti-neoplastic Efficacy Of Chemotherapeutic Agents And To Compare This Technique With The DNA Fragmentation Assay. Methods: Squamous Cell Carcinoma Cell Line HEp-2 Was Used In The Present Study. The Cytotoxic Drug Used In This Study Was Sesamol. Microscopic Slides Of Each Drug Concentration At Definite Post-treatment Durations Were Photomicrographed And Analyzed Using Image Analysis Software For The Estimation Of Nuclear Area Factor. Data Were Then Statistically Analyzed. To Assess Whether The Morphological Changes Induced By The Drug Depended On Activation Of The Apoptotic Pathway, It Was Investigated Whether It Could Induce Internucleosomal DNA Fragmentation.

Results: The Data Recorded Revealed A Decrease In The Mean Values Of NAF Of HEp-2 Cells Treated With Different Concentrations Of Sesamol 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 Sesamol Over Different Durations Revealed Absence Of DNA Laddering That Indicated DNA Fragmentation With 6.25 ?M Sesamol Concentration After 24 Hours Of Incubation. Conclusion: The Present Data Suggested That Estimation Of Nuclear Area Factor Is A Sensitive Predictor Of Early Apoptotic Effect Of Anti-cancer Therapy. DNA Fragmentation Assay Is A Useful Tool For Detection Of Late Apoptotic Changes That Discriminates Apoptotic Effects Of Anti-cancer Drugs From Necrotic Ones.

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