Journal List > Korean J Obstet Gynecol > v.53(5) > 1006411

Park, Kwack, Ryu, Lee, and Han: Measurement of apoptosis using M30 in culture media of cell lines treated with anti-cancer agents

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.

Figures and Tables

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).
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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).
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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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Table 1
Frequency of apoptosis in HeLa and OVCAR-3 cells assessed by M30-FITC immunofluorescence reactivity
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Values are expressed as the means±standard deviations (SD), the medium and interquartile range (IQR).

FITC: fluoresein isothiocyanate.

Table 2
Frequency of apoptosis in HeLa and OVCAR-3 cells assessed by M30-FITC immunofluorescence
kjog-53-401-i002

Values are expressed as the median and interquartile range (IQR).

FITC: fluoresein isothiocyanate.

*Comparison between control and chemo-treated group, Wilcoxon rank-sum (Mann-Whitney) test.

Comparison between HeLa and OVCAR-3, Wilcoxon rank-sum (Mann-Whitney) test.

Table 3
Measurement of apoptosis in media culturing HeLa and OVCAR-3 cells by the M30-ELISA
kjog-53-401-i003

Values are expressed as the median and interquartile range (IQR).

*Comparison between control and chemo-treated group, Wilcoxon rank-sum (Mann-Whitney) test.

Comparison between HeLa and OVCAR-3, Wilcoxon rank-sum (Mann-Whitney) test.

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