Expression of TUBB4B in mouse primary spermatocyte GC-2 cells and its regulatory effect on NF-κB and MAPK signaling pathway
OBJECTIVE: To explore the interaction between Tubulin beta 4B class IVb (TUBB4B) and Agtpbp1/cytosolic carboxypeptidase- like1 (CCP1) in mouse primary spermatocytes (GC-2 cells) and the role of TUBB4B in regulating the development of GC-2 cells.
METHODS: Lentiviral vectors were used to infect GC-2 cells to construct TUBB4B knockdown and negative control (NC-KD) cells. The stable cell lines with TUBB4B overexpression (Tubb4b-OE) and the negative control (NC-OE) cells were screened using purinomycin. RT-qPCR and Western blotting were used to verify successful cell modeling and explore the relationship between TUBB4B and CCP1 expressions in GC-2 cells. The effects of TUBB4B silencing and overexpression on the proliferation and cell cycle of GC-2 cells were evaluated using CCK8 assay and flow cytometry. The signaling pathway proteins showing significant changes in response to TUBB4B silencing or overexpression were identified using Western blotting and immunofluorescence assay and then labeled for verification at the cellular level.
RESULTS: Both TUBB4B silencing and overexpression in GC-2 cells caused consistent changes in the mRNA and protein expressions of CCP1 (P < 0.05). Similarly, TUBB4B expression also showed consistent changes at the mRNA and protein after CCP1 knockdown and restoration (P < 0.05). TUBB4B knockdown and overexpression had no significant effect on proliferation rate or cell cycle of GC-2 cells, but caused significant changes in the key proteins of the nuclear factor kappa-B (NF-κB) signaling pathway (p65 and p-p65) and the mitogen-activated protein kinase (MAPK) signaling pathway (ErK1/2 and p-Erk1/2) (P < 0.05); CCP1 knockdown induced significant changes in PolyE expression in GC-2 cells (P < 0.05).
CONCLUSIONS: TUBB4B and CCP1 interact via a mutual positive regulation mechanism in GC-2 cells. CCP-1 can deglutamize TUBB4B, and the latter is involved in the regulation of NF-κB and MAPK signaling pathways in primary spermatocytes.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:43 |
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| Enthalten in: |
Nan fang yi ke da xue xue bao = Journal of Southern Medical University - 43(2023), 6 vom: 20. Juni, Seite 1002-1009 |
| Sprache: |
Chinesisch |
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| Beteiligte Personen: |
Liu, T [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 17.07.2023 Date Revised 18.07.2023 published: Print Citation Status MEDLINE |
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| doi: |
10.12122/j.issn.1673-4254.2023.06.16 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM359399746 |
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| 024 | 7 | |a 10.12122/j.issn.1673-4254.2023.06.16 |2 doi | |
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| 100 | 1 | |a Liu, T |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Expression of TUBB4B in mouse primary spermatocyte GC-2 cells and its regulatory effect on NF-κB and MAPK signaling pathway |
| 264 | 1 | |c 2023 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 17.07.2023 | ||
| 500 | |a Date Revised 18.07.2023 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a OBJECTIVE: To explore the interaction between Tubulin beta 4B class IVb (TUBB4B) and Agtpbp1/cytosolic carboxypeptidase- like1 (CCP1) in mouse primary spermatocytes (GC-2 cells) and the role of TUBB4B in regulating the development of GC-2 cells | ||
| 520 | |a METHODS: Lentiviral vectors were used to infect GC-2 cells to construct TUBB4B knockdown and negative control (NC-KD) cells. The stable cell lines with TUBB4B overexpression (Tubb4b-OE) and the negative control (NC-OE) cells were screened using purinomycin. RT-qPCR and Western blotting were used to verify successful cell modeling and explore the relationship between TUBB4B and CCP1 expressions in GC-2 cells. The effects of TUBB4B silencing and overexpression on the proliferation and cell cycle of GC-2 cells were evaluated using CCK8 assay and flow cytometry. The signaling pathway proteins showing significant changes in response to TUBB4B silencing or overexpression were identified using Western blotting and immunofluorescence assay and then labeled for verification at the cellular level | ||
| 520 | |a RESULTS: Both TUBB4B silencing and overexpression in GC-2 cells caused consistent changes in the mRNA and protein expressions of CCP1 (P < 0.05). Similarly, TUBB4B expression also showed consistent changes at the mRNA and protein after CCP1 knockdown and restoration (P < 0.05). TUBB4B knockdown and overexpression had no significant effect on proliferation rate or cell cycle of GC-2 cells, but caused significant changes in the key proteins of the nuclear factor kappa-B (NF-κB) signaling pathway (p65 and p-p65) and the mitogen-activated protein kinase (MAPK) signaling pathway (ErK1/2 and p-Erk1/2) (P < 0.05); CCP1 knockdown induced significant changes in PolyE expression in GC-2 cells (P < 0.05) | ||
| 520 | |a CONCLUSIONS: TUBB4B and CCP1 interact via a mutual positive regulation mechanism in GC-2 cells. CCP-1 can deglutamize TUBB4B, and the latter is involved in the regulation of NF-κB and MAPK signaling pathways in primary spermatocytes | ||
| 650 | 4 | |a English Abstract | |
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a TUBB4B | |
| 650 | 4 | |a cytosolic carboxypeptidase-like1 | |
| 650 | 4 | |a male sterility | |
| 650 | 4 | |a mitogen-activated protein kinase MAPK pathway | |
| 650 | 4 | |a nuclear factor kappa-B pathway | |
| 650 | 4 | |a primary spermatocytes | |
| 650 | 7 | |a Agtpbp1 protein, mouse |2 NLM | |
| 650 | 7 | |a EC 3.4.17.- |2 NLM | |
| 650 | 7 | |a GTP-Binding Proteins |2 NLM | |
| 650 | 7 | |a EC 3.6.1.- |2 NLM | |
| 650 | 7 | |a Mitogen-Activated Protein Kinases |2 NLM | |
| 650 | 7 | |a EC 2.7.11.24 |2 NLM | |
| 650 | 7 | |a NF-kappa B |2 NLM | |
| 650 | 7 | |a RNA, Messenger |2 NLM | |
| 650 | 7 | |a Serine-Type D-Ala-D-Ala Carboxypeptidase |2 NLM | |
| 650 | 7 | |a EC 3.4.16.4 |2 NLM | |
| 650 | 7 | |a Tubulin |2 NLM | |
| 700 | 1 | |a Wang, W |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, T |e verfasserin |4 aut | |
| 700 | 1 | |a Liu, S |e verfasserin |4 aut | |
| 700 | 1 | |a Bian, Y |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, C |e verfasserin |4 aut | |
| 700 | 1 | |a Xiao, R |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Nan fang yi ke da xue xue bao = Journal of Southern Medical University |d 2006 |g 43(2023), 6 vom: 20. Juni, Seite 1002-1009 |w (DE-627)NLM16087646X |x 1673-4254 |7 nnns |
| 773 | 1 | 8 | |g volume:43 |g year:2023 |g number:6 |g day:20 |g month:06 |g pages:1002-1009 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.12122/j.issn.1673-4254.2023.06.16 |3 Volltext |
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