Artikel
Effects of protocadherin gamma C3 (PCDHGC3) knockout on proliferation, invasion and self-renewal abilities in glioblastoma cell lines
Auswirkungen von Protocadherin-gamma-C3 (PCDHGC3)-Knockout auf die Proliferations-, Invasions- und Selbsterneuerungsfähigkeiten von Glioblastomzelllinien
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Autoren
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Christoph D. Kempe
- Universität Würzburg, Klinik und Poliklinik für Anästhesiologie, Würzburg, Deutschland
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Francisco Baptista
- Universität Würzburg, Klinik und Poliklinik für Anästhesiologie, Würzburg, Deutschland
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Maria C. Fabregat
- Universität Würzburg, Klinik und Poliklinik für Anästhesiologie, Würzburg, Deutschland
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Nuo Li
- Universität Würzburg, Klinik und Poliklinik für Anästhesiologie, Würzburg, Deutschland
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Carsten Hagemann
- Universität Würzburg, Neurochirurgische Klinik und Poliklinik, Würzburg, Deutschland
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Malgorzata Burek
- Universität Würzburg, Klinik und Poliklinik für Anästhesiologie, Würzburg, Deutschland
| Veröffentlicht: | 8. Mai 2019 |
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Gliederung
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Objective: Protocadherins belong to the cadherin family of adhesions molecules. They are highly expressed in the central nervous system. It has been shown that several Protocadherins function as tumour suppressors. A member of this protein family, Protocadherin gamma C3 (PCDHGC3) is strongly expressed in glioblastoma (GBM) and has been previously shown to down-regulate the mTOR signalling pathway. Therefore, we investigated the effects of PCDHGC3 knockout in GBM cell lines on survival, migration and invasion.
Methods: We tested the expression of PCDHGC3 in the GBM cell lines GaMG, U87 MG, U138 MG and U343 MG by Western blotting (WB) with specific anti-PCDHGC3 antibodies. The cell lines with the strongest PCDHGC3 expression were selected for genome editing using the CRISPR/Cas Method and successful knockout was confirmed by WB and qPCR. The growth rate was measured using MTT-assay. Moreover, proliferation, migration, matrigel invasion, colony formation ability as well as responsiveness to the standard GBM therapy with temozolomide were compared between wild type and knockout cells.
Results: Of the examined GBM cell lines, U138 MG and U343 MG had the highest PCDHGC3 expression. A complete deletion of PCDHGC3 was achieved in U138 MG and U343 MG. Compared to the wild type cells, the PCDHGC3 knockout led to a 1.7±0.12 fold increased growth rate (p<0.0001, unpaired t test) of U138 MG cells. In contrast, the PCDHGC3 knockout in U343 MG cells showed a 1.5±0.26 fold decreased growth rate (p< 0.0001, unpaired t test). Moreover, the PCDHGC3 knockout cells were more sensitive to temozolomide.
Conclusion: PCDHGC3 seems to be one of the key players in GBM cell physiology. This may be due to its involvement in signalling pathways such as mTOR. The latter has already become an important therapeutic target for the treatment of GBM. Thus, influencing PCDHGC3 expression could further improve this already promising therapeutic strategy by inhibiting tumour growth and enhancing the tumour responsiveness to the standard therapy.
