Revealing the Role of Alternariol in the Local Steroidogenesis in Human Prostate Normal and Cancer Cells
Abstract
:1. Introduction
2. Results
2.1. AOH Modulates the Viability of Prostate Normal and Cancer Cell Lines in a Dose- and Time-Dependent Manner
2.2. AOH Modulates the Local Steroidogenesis in Prostate Normal and Cancer Cells in the Presence of DHEA
2.3. AOH Affects the Expression of Caveolin-1 (CAV-1) in Prostate Cells
2.4. AOH Modulates Cell Cycle and Apoptosis in Prostate Cells
3. Discussion
4. Materials and Methods
4.1. Cell Culture and Experimental Treatments
4.2. Cell Viability Assay
4.3. Annexin V Staining Assay
4.4. Cell Cycle
4.5. Steroid Assays
4.6. RNA Extraction and Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR)
4.7. Western Blot
4.8. Immunohistochemistry Staining and Confocal Microscopy
4.9. Statistical Analysis
Author Contributions
Funding
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Cell Line | Treatment | CAV-1 |
---|---|---|
PNT1A | 10 μM AOH | 0.62 |
0.1 μM AOH | 1.06 | |
100 nM DHEA | 1.08 | |
10 μM AOH + 100 nM DHEA | 1.61 | |
0.1 μM AOH + 100 nM DHEA | 1.48 | |
Cnt | 1.00 | |
PC3 | 10 μM AOH | 0.67 |
0.1 μM AOH | 0.44 | |
100 nM DHEA | 0.95 | |
10 μM AOH + 100 nM DHEA | 0.62 | |
0.1 μM AOH + 100 nM DHEA | 0.57 | |
Cnt | 1.00 |
Cell Line | Treatment | Casp3 | Cleaved Casp3 |
---|---|---|---|
PNT1A | 10 μM AOH | 0.94 | 0.70 |
0.1 μM AOH | 0.89 | 0.66 | |
100 nM DHEA | 1.33 | 1.03 | |
10 μM AOH + 100 nM DHEA | 0.92 | 0.71 | |
0.1 μM AOH + 100 nM DHEA | 1.31 | 0.80 | |
Cnt | 1.00 | 1.00 | |
PC3 | 10 μM AOH | 2.15 | 1.83 |
0.1 μM AOH | 1.07 | 1.42 | |
100 nM DHEA | 1.51 | 1.92 | |
10 μM AOH + 100 nM DHEA | 0.98 | 0.98 | |
0.1 μM AOH + 100 nM DHEA | 0.99 | 1.38 | |
Cnt | 1.00 | 1.00 |
Gene | Sequence (5′-3′) | Product Size (bp) |
---|---|---|
CYP11A1 | For CCAGAACGATTCCTCATCC | 126 |
Rev CATCACCTCCTGGTTCAG | ||
CYP17A1 | For GAAGTTATCATCAATCTGTGGG | 119 |
Rev ACTGACGGTGAGATGAGC | ||
Rev AAGATGTCTGGTTTGATGAGGAG | ||
HSD3B2 | For CTTGGTGTCACTCACAGAGAG | 128 |
Rev GTAGATGAAGACTGGCACACTG | ||
Rev CACCTCCAATTGTGACATAA | ||
STAR | For CATGGAGAGGCTCTATGAAGA | 128 |
Rev CAGCCAGCTCGTGAGTAAT | ||
ANXA5 | For ACCCTCTCGGCTTTATGATGCT | 116 |
Rev TGGCTCTCAGTTCTTCAGGTGT | ||
Rev TGCTGGACAGAAATGTGTACACTCCAGA | ||
ESR2 | For ACACCTGGGCACCTTTCTCCTTTA | 90 |
Rev TCTTGCTTCACACCAGGGACTCTT | ||
AR | For GGGAGGTTACACCAAAGGGC | 102 |
Rev AGAGACAGGGTAGACGGCAG | ||
CAV-1 | Rev GAACTTGAAATTGGCACCAGG | 139 |
For ACCCACTCTTTGAAGCTGTTG | ||
CDK1N1A | Rev CTGAGACTAAGGCAGAAGATGT | 133 |
For GACAGATTTCTACCACTCCAA | ||
CDK2 | Rev GAGCAGAGGCATCCATGAATT | 126 |
For TGCTTAAGGAGCTTAACCATCC | ||
TP53 | Rev TTTATGGCGGGAGGTAGA | 102 |
For TTGGAACTCAAGGATGCC | ||
For GCAAGACTGTTAGCCCTCAA | ||
RPLP0 | For ACGGATTACACCTTCCCACTTGCTAAAAGGTC | 69 |
Rev AGCCACAAAGGCAGATGGATCAGCCAAG | ||
RPS17 | For AAGCGCGTGTGCGAGGAGATCG | 87 |
Rev TCGCTTCATCAGAT GCGTGACATAACCTG | ||
H3F3A | For AGGACTTTAAAAGATCTGCGCTTCCAGAG | 74 |
Rev ACCAGATAGGCCTCACTTGCCTCCTGC |
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Urbanek, K.A.; Kowalska, K.; Habrowska-Górczyńska, D.E.; Kozieł, M.J.; Domińska, K.; Piastowska-Ciesielska, A.W. Revealing the Role of Alternariol in the Local Steroidogenesis in Human Prostate Normal and Cancer Cells. Int. J. Mol. Sci. 2023, 24, 9513. https://doi.org/10.3390/ijms24119513
Urbanek KA, Kowalska K, Habrowska-Górczyńska DE, Kozieł MJ, Domińska K, Piastowska-Ciesielska AW. Revealing the Role of Alternariol in the Local Steroidogenesis in Human Prostate Normal and Cancer Cells. International Journal of Molecular Sciences. 2023; 24(11):9513. https://doi.org/10.3390/ijms24119513
Chicago/Turabian StyleUrbanek, Kinga Anna, Karolina Kowalska, Dominika Ewa Habrowska-Górczyńska, Marta Justyna Kozieł, Kamila Domińska, and Agnieszka Wanda Piastowska-Ciesielska. 2023. "Revealing the Role of Alternariol in the Local Steroidogenesis in Human Prostate Normal and Cancer Cells" International Journal of Molecular Sciences 24, no. 11: 9513. https://doi.org/10.3390/ijms24119513