Korean J Obstet Gynecol.  2010 May;53(5):401-409. 10.5468/kjog.2010.53.5.401.

Measurement of apoptosis using M30 in culture media of cell lines treated with anti-cancer agents

• Park SH
• Kwack HS
• Ryu KS
• Lee Y
• Han KT

• KMID: 2273871
• DOI: http://doi.org/10.5468/kjog.2010.53.5.401

Abstract

OBJECTIVE
We investigated a possible use of the induced apoptosis as a biomarker in the cells and their media treated with commonly used anti-cancer agents in gynecologic malignancies.
METHODS
After treatments with low and high concentrations of paclitaxel, cisplatin, and camptothecin in HeLa and OVCAR-3 cells, the levels of M30 antigen were detected in the cells and their media by immunofluorescence staining and ELISA methods, respectively.
RESULTS
The percentages of M30-fluoresein isothiocyanate (FITC) positive cells in HeLa and OVCAR-3 cells treated with paclitaxel, cisplatin, and camptothecin were 4.3% vs 18.1% vs 34.87% and 4.07% vs 18.6% vs 32.63%, 4.3% vs 17.87% vs 32.38% and 4.07% vs 16.83% vs 32%, and 4.3% vs 16.75% vs 31.3% and 4.07% vs 15.18% vs 29.9% in control, low dose, and hight dose groups, respectively (P<0.001). M30 antigen levels (U/L) measured in culture media of HeLa and OVCAR-3 cells treated with paclitaxel, cisplatin, and camptothecin were 53.03 vs 101.53 vs 355.59 and 86 vs 114.41 vs 412.04, 53.03 vs 79.84 vs 327.64 and 86 vs 125.44 vs 385.09, and 53.03 vs 88.41 vs 295.005 and 86 vs 108.42 vs 263.1 in control, low dose, and hight dose groups, respectively (P<0.001).
CONCLUSION
Our results obtained in this preclinical study suggests that measurement of the levels of M30 antigen may help to predict the clinical responses and to select the effective anti-cancer agents in clinical settings, rapidly and quantitatively.

Keyword

Apoptosis; M30 antigen; HeLa cells; OVCAR-3

MeSH Terms

Apoptosis
Camptothecin
Cell Line
Cisplatin
Culture Media
Enzyme-Linked Immunosorbent Assay
Fluorescent Antibody Technique
HeLa Cells
Humans
Isothiocyanates
Paclitaxel
Camptothecin
Cisplatin
Culture Media
Isothiocyanates
Paclitaxel

Figure

• 

  Fig. 1 Microscopic findings of M30-FITC immunofluorescence in HeLa and OVCAR-3 cells according to each of concentrations of paclitaxel, cisplatin, and camptothecin after culturing for 24 hours (×200).

• 

  Fig. 2 Percentages of cells with positive immunofluorescence after staining by the M30-FITC monoclonal antibody in HeLa (A) and OVCAR-3 (B) cells according to each of concentrations of paclitaxel, cisplatin, and camptothecin after culturing for 24 hours. Shown are the median and interquartile range (IQR).

• 

  Fig. 3 Increase in the levels of M30-antigen measuring by M30 ELISA in media culturing HeLa (A) and OVCAR-3 (B) cells according to each of concentrations of paclitaxel, cisplatin, and camptothecin after culturing for 24 hours. Shown are the median and interquartile range (IQR).

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