PTPRK Expression Is Downregulated in Drug Resistant Ovarian Cancer Cell Lines, and Especially in ALDH1A1 Positive CSCs-Like Populations
Abstract
:1. Introduction
2. Results
2.1. Analyses of PTPRK Gene Expression in Drug-Resistant Ovarian Cancer Cell Lines
2.2. Immunofluorescence Analysis of PTPRK and pTYR in Drug Sensitive and Resistant Cell Lines
2.3. Western Blot Analysis of PTPRK and pTYR
2.4. Early Response to Topotecan (TOP)-Treatment in Ovarian Cancer Cell Lines
2.5. Co-Expression of PTPRK and ALDH1A1
2.6. Immunohistochemistry
3. Discussion
4. Materials and Methods
4.1. Reagents and Antibodies
4.2. Cell Lines and Cell Culture
4.3. Examination of Gene Expression Using Q-PCR
4.4. Protein Isolation from Cell Culture
4.5. SDS-PAGE and Western Blot Analysis of PTPRK and p-Tyr
4.6. Immunofluorescence Analysis
4.7. Double Immunofluorescence Analysis
4.8. Incubation of Cells with TOP
4.9. Immunohistochemistry
4.10. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
Abbreviations
PTPRK | Protein Tyrosine Phosphatase Receptor Type K |
CSCs | Cancer Stem Cells |
CR | Cisplatin Resistance |
DR | Doxorubicin Resistance |
MR | Methotrexate Resistance |
PR | Paclitaxel Resistance |
TR | Topotecan Resistance |
VR | Vincristine Resistance |
ALDH1A1 | Aldehyde Dehydrogenase 1 Family Member A1 |
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Transcript | Sequence (5′–3′ Direction) | Ensembl Transcript (ENST) Number (Available Online: http://www.ensembl.org) | Product Size (bp) |
---|---|---|---|
PTPRK | CCCAGGACCTCCACTAATCA ATTCCCAGTCCACAGCAATC | 00000368226 | 110 bp |
GADPH | GAAGGTGAAGGTCGGAGTCA GACAAGCTTCCCGTTCTCAG | 00000229239 | 199 bp |
β-actin | TCTGGCACCACACCTTCTAC GATAGCACAGCCTGGATAGC | 00000331789 | 169 bp |
HRPT1 | CTGAGGATTTGGAAAGGGTG AATCCAGCAGGTCAGCAAAG | 00000298556 | 156 bp |
β2M | CGCTACTCTCTCTTTCTGGC ATGTCGGATGGATGAAACCC | 00000558401 | 133 bp |
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Świerczewska, M.; Sterzyńska, K.; Wojtowicz, K.; Kaźmierczak, D.; Iżycki, D.; Nowicki, M.; Zabel, M.; Januchowski, R. PTPRK Expression Is Downregulated in Drug Resistant Ovarian Cancer Cell Lines, and Especially in ALDH1A1 Positive CSCs-Like Populations. Int. J. Mol. Sci. 2019, 20, 2053. https://doi.org/10.3390/ijms20082053
Świerczewska M, Sterzyńska K, Wojtowicz K, Kaźmierczak D, Iżycki D, Nowicki M, Zabel M, Januchowski R. PTPRK Expression Is Downregulated in Drug Resistant Ovarian Cancer Cell Lines, and Especially in ALDH1A1 Positive CSCs-Like Populations. International Journal of Molecular Sciences. 2019; 20(8):2053. https://doi.org/10.3390/ijms20082053
Chicago/Turabian StyleŚwierczewska, Monika, Karolina Sterzyńska, Karolina Wojtowicz, Dominika Kaźmierczak, Dariusz Iżycki, Michał Nowicki, Maciej Zabel, and Radosław Januchowski. 2019. "PTPRK Expression Is Downregulated in Drug Resistant Ovarian Cancer Cell Lines, and Especially in ALDH1A1 Positive CSCs-Like Populations" International Journal of Molecular Sciences 20, no. 8: 2053. https://doi.org/10.3390/ijms20082053