Effect of Interaction between Chromium(VI) with 17β-Estradiol and Its Metabolites on Breast Cancer Cell Lines MCF-7/WT and MDA-MB-175-VII: Preliminary Study
Abstract
:1. Introduction
2. Results
2.1. Viability Assay of Single Compounds (First Stage, First Model)
2.1.1. Effect of 17β-E2, Its Metabolites, and Cr(VI) on MCF-7/WT Viability
2.1.2. Effect of 17β-E2, Its Metabolites, and Cr(VI) on MDA-MB-175-VII Viability
2.2. Viability Assay of Combined Action of Estrogens with Cr(VI) (First Stage, Second Model)
2.2.1. Effect of Combined Action of 17β-E2 with Cr(VI) on MCF-7/WT Viability
2.2.2. Effect of Combined Action of 2-MeOE2 with Cr(VI) on MCF-7/WT Viability
2.2.3. Effect of Combined Action of 4-OHE2 with Cr(VI) on MCF-7/WT Viability
2.2.4. Effect of Combined Action of 16α-OHE1 with Cr(VI) on MCF-7/WT Viability
2.2.5. Effect of Combined Action of 17β-E2 with Cr(VI) on MDA-MB-175-VII Viability
2.2.6. Effect of Combined Action of 17β-E2 Metabolites with Cr(VI) on MDA-MB-175-VII Viability
2.3. Genotoxicity of Single Compounds (Second Stage, First Model: Single Action) Genotoxicity of 17β-E2 and Its Metabolites
Genotoxicity of Cr(VI)
2.4. Genotoxicity of Combined Action of Estrogens with Cr(VI) (Second Stage, Second Model)
2.5. SOD1 Estimation: Immunocytochemical Staining
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Compounds
4.3. MTT Assay
4.4. Neutral Comet Assay (NCA)
4.5. SOD1 Estimation: Immunocytochemical Staining
4.6. Schematic Model for Viability and Genotoxicity Testing
4.7. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
References
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IC 50 for Examined Compounds | |||
---|---|---|---|
Cr(VI) | 0.001 µM 17β-E2 | 10 µM 17β-E2 | 25 µM 17β-E2 |
5.2251 | 4.2423 | 2.1506 | 0.4278 |
Cr(VI) | 0.001 µM2-MeOE2 | 0.5 µM 2-MeOE2 | 10 µM 2-MeOE2 |
5.2251 | 6.4528 | 3.2515 | 2.1813 |
Cr(VI) | 0.001 µM 4-OHE2 | 0.5 µM 4-OHE2 | 10 µM 4-OHE2 |
5.2251 | 3.2589 | 4.5478 | 6.8237 |
Cr(VI) | 0.001 µM 16α-OHE1 | 10 µM16α-OHE1 | 25 µM16α-OHE1 |
5.2251 | 4.2465 | 3.5783 | 6.8957 |
IC 50 for Examined Compounds | ||||
---|---|---|---|---|
17β-E2 | Cr(VI) | 7 days pre-inc. 0.001 µM 17β-E2 | 7 days pre-inc. 10 µM 17β-E2 | 7 days pre-inc. 25 µM 17β-E2 |
5.2251 | 6.6895 | 4.9685 | 12.3467 | |
2-MeOE2 | Cr(VI) | 7 days pre-inc. 0.001 µM 2-MeOE2 | 7 days pre-inc. 0.5 µM 2-MeOE2 | days pre-inc. 10 µM 2-MeOE2 |
5.2251 | 4.2584 | 6.9854 | 0.2584 | |
4-OHE2 | Cr(VI) | 7 days pre-inc. 0.001 µM 4-OHE2 | 7 days pre-inc. 0.5 µM 4-OHE2 | 7 days pre-inc. 10 µM 4-OHE2 |
5.2251 | 3.8255 | 4.2857 | 7.2824 | |
16α-OHE1 | Cr(VI) | 7 days pre-inc. 0.001µM 16α-OHE1 | 7 days pre-inc. 10 µM 16α-OHE1 | 7 days pre-inc. 25 µM16α-OHE1 |
5.2251 | 7.2867 | 9.5857 | 2.9487 |
IC 50 for Examined Compounds | |||
---|---|---|---|
Cr(VI) | 0.001 µM 17β-E2 | 10 µM 17β-E2 | 25 µM 17β-E2 |
5.2251 | 3.9223 | 2.1789 | 0.6267 |
Cr(VI) | 0.001 µM2-MeOE2 | 0.5 µM 2-MeOE2 | 10 µM 2-MeOE2 |
5.2251 | 5.8728 | 3.9875 | 1.9874 |
Cr(VI) | 0.001 µM 4-OHE2 | 0.5 µM 4-OHE2 | 10 µM 4-OHE2 |
5.2251 | 2.9749 | 3.7838 | 7.4537 |
Cr(VI) | 0.001 µM 16α-OHE1 | 10 µM16α-OHE1 | 25 µM16α-OHE1 |
5.2251 | 3.9865 | 2.9783 | 6.0743 |
IC 50 for Examined Compounds | ||||
---|---|---|---|---|
17β-E2 | Cr(VI) | 7 days pre-inc. 0.001 µM 17β-E2 | 7 days pre-inc. 10 µM 17β-E2 | 7 days pre-inc. 25 µM 17β-E2 |
4.5671 | 5.9795 | 5.3485 | 10.8767 | |
2-MeOE2 | Cr(VI) | 7 days pre-inc. 0.001 µM 2-MeOE2 | 7 days pre-inc. 0.5 µM 2-MeOE2 | 7 days pre-inc. 10 µM 2-MeOE2 |
6.1151 | 3.9985 | 7.1540 | 0.1875 | |
4-OHE2 | Cr(VI) | 7 days pre-inc. 0.001 µM 4-OHE2 | 7 days pre-inc. 0.5 µM 4-OHE2 | 7 days pre-inc. 10 µM 4-OHE2 |
5.5461 | 4.1563 | 4.7854 | 6.8374 | |
16α-OHE1 | Cr(VI) | 7 days pre-inc. 0.001µM 16α-OHE1 | 7 days pre-inc. 10 µM16α-OHE1 | 7 days pre-inc. 25 µM16α-OHE1 |
4.8451 | 6.1127 | 7.9987 | 2.3567 |
Compound | % of Stained Cells | Intensity of the Reaction |
---|---|---|
17β-E2 [µM] | ||
0.001 | 42% | +++ |
0.1 | 59% | ++ |
10 | 56% | ++/+++ |
25 | 45% | ++/+++ |
Cr(VI)[µM] | ||
0.1 | 29% | +/++ |
1 | 32% | ++ |
5 | 22% | ++ |
10 | 37% | ++ |
50 | 18% | ++ |
2ME2 [µM] | ||
0.001 | 38% | ++ |
0.1 | 42% | ++ |
0.5 | 39% | ++ |
10 | 27% | ++ |
4-OH [µM] | ||
0.001 | 25% | ++ |
0.5 | 18% | ++ |
10 | 14% | ++ |
25 | 34% | ++/+++ |
16α-OHE1 [µM] | ||
0.001 | 28% | ++ |
1 | 32% | ++ |
10 | 19% | ++ |
25 | 35% | ++ |
Control | 43% | ++/+++ |
Cr(VI) [µM] | % of Stained Cells | Intensity of the Reaction | Cr(VI) [µM] | % of Stained Cells | Intensity of the Reaction |
---|---|---|---|---|---|
17β-E2 0.001 µM | 17β-E2 10 µM | ||||
0.1 | 21% | ++ | 0.1 | 49% | ++ |
1 | 39% | +/++ | 1 | 41% | ++ |
5 | 28% | ++ | 5 | 38% | ++ |
10 | 27% | ++/+++ | 10 | 46% | ++/+++ |
50 | 52% | ++/+++ | 50 | 68% | +++ |
17β-E2 0.1 µM | 17β-E2 25 µM | ||||
0.1 | 32% | ++/+++ | 0.1 | 47% | ++ |
1 | 38% | ++/+++ | 1 | 38% | ++ |
5 | 41% | ++ | 5 | 39% | ++ |
10 | 49% | ++ | 10 | 37% | ++ |
50 | 43% | ++ | 50 | 24% | ++ |
Control | 43% | ++/+++ | Control | 43% | ++/+++ |
Cr(VI) [µM] | % of Stained Cells | Intensity of the Reaction | Cr(VI) [µM] | % of Stained Cells | Intensity of the Reaction |
---|---|---|---|---|---|
2-MeOE2 0.001 µM | 2-MeOE2 0.5 µM | ||||
0.1 | 42% | ++ | 0.1 | 54% | ++ |
1 | 56% | ++ | 1 | 44% | ++ |
5 | 61% | ++/+++ | 5 | 27% | ++ |
10 | 70% | ++/+++ | 10 | 42% | ++ |
50 | 76% | ++/+++ | 50 | 51% | ++ |
2-MeOE2 0.1 µM | 2-MeOE2 10 µM | ||||
0.1 | 60% | ++ | 0.1 | 45% | ++ |
1 | 50% | ++ | 1 | 43% | ++ |
5 | 45% | ++ | 5 | 35% | + |
10 | 30% | ++/+++ | 10 | 43% | ++ |
50 | 50% | ++/+++ | 50 | 59% | ++ |
Control | 43% | ++/+++ | Control | 43% | ++/+++ |
Cr(VI) [µM] | % of Stained Cells | Intensity of the Reaction | Cr(VI) [µM] | % of Stained Cells | Intensity of the Reaction |
---|---|---|---|---|---|
4-OHE2 0.001 µM | 4-OHE2 0.5 µM | ||||
0.1 | 42% | +++ | 0.1 | 22% | ++ |
1 | 36% | ++ | 1 | 32% | ++ |
5 | 47% | ++ | 5 | 17% | ++ |
10 | 40% | ++ | 10 | 22% | +/++ |
50 | 56% | +++ | 50 | 21% | ++ |
4-OHE2 10 µM | 4-OHE2 25 µM | ||||
0.1 | 25% | ++ | 0.1 | 25% | ++ |
1 | 16% | ++ | 1 | 34% | ++ |
5 | 27% | ++ | 5 | 21% | + |
10 | 23% | ++ | 10 | 29% | +/++ |
50 | 37% | ++ | 50 | 28% | ++ |
Control | 43% | ++/+++ | Control | 43% | ++/+++ |
Cr(VI) [µM] | % of Stained Cells | Intensity of the Reaction | Cr(VI)2 [µM] | % of Stained Cells | Intensity of the Reaction |
---|---|---|---|---|---|
16α-OHE1 0.001 µM | 16α-OHE1 10 µM | ||||
0.1 | 35% | ++ | 0.1 | 43% | ++ |
1 | 29% | ++ | 1 | 44% | ++ |
5 | 45% | ++ | 5 | 38% | ++ |
10 | 28% | ++ | 10 | 56% | + |
50 | 35% | + | 50 | 34% | + |
16α-OHE1 1 µM | 16α-OHE1 25 µM | ||||
0.1 | 50% | ++ | 0.1 | 45% | ++ |
1 | 42% | ++ | 1 | 33% | ++ |
5 | 39% | ++ | 5 | 45% | ++ |
10 | 49% | +/++ | 10 | 53% | ++ |
50 | 38% | ++ | 50 | 59% | ++ |
Control | 43% | ++/+++ | Control | 43% | ++/+++ |
Name of Compound | Structure and Manufacturer’s Name | Molecular Mass of Compound |
---|---|---|
17β-E2 | SIGMA-ALDRICH | 272.38 g/mol |
2-MeOE2 | SIGMA-ALDRICH | 302.41 g/mol |
4-OHE2 | SIGMA-ALDRICH | 288.38 g/mol |
16α-OHE1 | STERALOIDS | 286.40 g/mol |
potassium chromate (VI) | K2CrO4 SIGMA-ALDRICH | 194.19 g/mol |
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Sawicka, E.; Kulbacka, J.; Drąg-Zalesińska, M.; Woźniak, A.; Piwowar, A. Effect of Interaction between Chromium(VI) with 17β-Estradiol and Its Metabolites on Breast Cancer Cell Lines MCF-7/WT and MDA-MB-175-VII: Preliminary Study. Molecules 2023, 28, 2752. https://doi.org/10.3390/molecules28062752
Sawicka E, Kulbacka J, Drąg-Zalesińska M, Woźniak A, Piwowar A. Effect of Interaction between Chromium(VI) with 17β-Estradiol and Its Metabolites on Breast Cancer Cell Lines MCF-7/WT and MDA-MB-175-VII: Preliminary Study. Molecules. 2023; 28(6):2752. https://doi.org/10.3390/molecules28062752
Chicago/Turabian StyleSawicka, Ewa, Julita Kulbacka, Małgorzata Drąg-Zalesińska, Arkadiusz Woźniak, and Agnieszka Piwowar. 2023. "Effect of Interaction between Chromium(VI) with 17β-Estradiol and Its Metabolites on Breast Cancer Cell Lines MCF-7/WT and MDA-MB-175-VII: Preliminary Study" Molecules 28, no. 6: 2752. https://doi.org/10.3390/molecules28062752