Artikel
The correlation of Vimentin3 and Toll-like receptor 4 in prostate cancer cell lines
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Autoren
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Jasmin Behring
- Uniklinik Köln, Klinik für Urologie, Köln, Germany
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Barbara Köditz
- Uniklinik Köln, Klinik für Urologie, Köln, Germany
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David Pfister
- Uniklinik Köln, Klinik für Urologie, Köln, Germany
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Axel Heidenreich
- Uniklinik Köln, Klinik für Urologie, Köln, Germany
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Melanie von Brandenstein
- Uniklinik Köln, Klinik für Urologie, Köln, Germany
| Veröffentlicht: | 28. März 2023 |
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Gliederung
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Introduction: The Toll-like receptor 4 (TLR4) is expressed in both benign and malignant tissue of the prostate. The long-term signalling of TLR4 may promote tumour survival and disease progression. Findings suggest that TLR4 levels correlate with the metastatic potential of prostate cancer (PCa). It is known that the truncated version of vimentin, Vim3, migrates into the nucleus with a yet unknown function. It was possible to identify, according to the unique C-terminal ending of Vim3, hypothetical DNA binding sites within the promotor region of TLR4.
Methods: PC3, DU145, LnCap and BPH cells were treated with ET-1 (Vim3 enhancer), WithA (Vimentin inhibitor) and LPS (TLR4 enhancer). A scratch assay followed by Western Blotting (WB) and ELISA was performed. With immunofluorescence (IF) the localization of the proteins was shown. To determine if Vim3 interacts with the hypothesized DNA region of TLR4, a pulldown assay was performed. siRNA knockdown of Vim3 was conducted in parallel with a scratch assay. 50 µl of patient serum was analysed via TLR4 ELISA.
Results: A significant upregulation of Vim3 and TLR4 could be detected via WB and ELISA. The upregulation of both proteins was highest in LPS treated cells. In scratch assays, the migration behavior significantly increased after ET-1 and LPS stimulation, while it decreased after WithA. In a Vim3 pulldown assay, interaction with TLR4 promotor region was shown. Vim3 and TLR4 are present in both the cytoplasm and the nucleus as shown via IF. After siRNA knockdown of Vim3 a decreased migration behaviour was observed. In serum ELISA a significant upregulation in PCa (n=60) was detectable compared with healthy control serum samples (n=15).
Conclusion: Here, we could show that Vim3 interacts with the promoter region of TLR4. Vim3 seems to be involved in the proliferation of prostate cancer cells. Concluding this, Vim3 seems to be an important TLR4 activator. The deeper biological impact of Vim3 in the development of PCa needs to be analysed in the future.
