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Use of the monoclonal antibody M30 for detecting HSG cell apoptosis
1Department of Oral Pathology, Meikai University School of Dentistry, Saitama, Japan. kusama@dent.meikai.ac.jp
Abstract:
An immunocytochemical method using a monoclonal antibody (MoAb), M30, which reacts with the product resulting from the cleavage of cytokeratin 18 by activated caspase, was applied to detect the apoptosis of human salivary gland tumor (HSG) cells induced by epigallocatechin gallate (EGCG), gallic acid (GA) and sodium ascorbate (SA). EGCG, GA and SA dose-dependently induced HSG cell death. Immunoreactive products were significantly observed in the cytoplasm of HSG cells after treatment with all these compounds. The reactions occurred with lower concentrations of these agents and after shorter treatment times, in comparison with DNA fragmentation detected by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method. These results suggest that immunocytochemical staining with the MoAb M30 may be useful for detecting the apoptosis-inducing activities of various chemical compounds.
Insights
Epigallocatechin gallate (EGCG), gallic acid (GA), and sodium ascorbate (SA) induce human salivary gland tumor (HSG) cell death. Immunocytochemical staining with M30 MoAb detects apoptosis earlier than DNA fragmentation methods.
Area of Science:
- Biochemistry
- Cell Biology
- Oncology
Background:
- Apoptosis, or programmed cell death, is crucial in regulating cell populations and preventing tumor development.
- Investigating novel therapeutic agents for human salivary gland tumors (HSG) is essential.
- Early detection of apoptosis is key to assessing treatment efficacy.
Purpose of the Study:
- To evaluate the apoptosis-inducing effects of epigallocatechin gallate (EGCG), gallic acid (GA), and sodium ascorbate (SA) on HSG cells.
- To assess the utility of a monoclonal antibody (MoAb) M30 for detecting early apoptotic events.
- To compare the sensitivity of MoAb M30 staining with the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay.
Main Methods:
- Human salivary gland tumor (HSG) cells were treated with varying concentrations of EGCG, GA, and SA.
- Immunocytochemistry using the M30 monoclonal antibody, which targets caspase-cleaved cytokeratin 18, was performed.
- DNA fragmentation was assessed using the TUNEL assay for comparison.
Main Results:
- EGCG, GA, and SA demonstrated dose-dependent induction of HSG cell death.
- Significant cytoplasmic immunoreactivity with MoAb M30 was observed in treated HSG cells.
- Apoptotic changes were detected earlier and at lower compound concentrations using MoAb M30 compared to the TUNEL method.
Conclusions:
- EGCG, GA, and SA possess apoptosis-inducing properties relevant to HSG cells.
- Immunocytochemical staining with MoAb M30 is a sensitive method for detecting early apoptosis.
- MoAb M30 staining offers a potentially more rapid and sensitive approach for evaluating apoptosis-inducing agents compared to traditional DNA fragmentation assays.