Cargando…
Activation of the mTOR pathway promotes neurite growth through upregulation of CD44 expression
OBJECTIVE: To explore the intrinsic mechanism of the mammalian target of rapamycin (mTOR) pathway activation and promotion of neuronal axon growth. METHODS: Human neuroblastoma cells, SH-SY5Y, were induced with all-trans retinoic acid (ATRA; 10 μM for three days) which differentiated the cell line i...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
SAGE Publications
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10288410/ https://www.ncbi.nlm.nih.gov/pubmed/37340717 http://dx.doi.org/10.1177/03000605231178510 |
_version_ | 1785062077157605376 |
---|---|
author | Zhang, Jiwei Gan, Wenjuan Peng, Ru Lu, Lei Lu, Weiqing Liu, Jiamei |
author_facet | Zhang, Jiwei Gan, Wenjuan Peng, Ru Lu, Lei Lu, Weiqing Liu, Jiamei |
author_sort | Zhang, Jiwei |
collection | PubMed |
description | OBJECTIVE: To explore the intrinsic mechanism of the mammalian target of rapamycin (mTOR) pathway activation and promotion of neuronal axon growth. METHODS: Human neuroblastoma cells, SH-SY5Y, were induced with all-trans retinoic acid (ATRA; 10 μM for three days) which differentiated the cell line into a neuronal-like state. Immunohistochemical staining was used to detect the differentiation status of the neuronal-like cells. Phosphatase and tensin homolog (PTEN) RNA interference (RNAi) experiments were performed on the differentiated cells; reverse transcription-polymerase chain reaction (RT-PCR) detected transcriptional levels of PTEN following 24 h of interference. After 36 h, western blot assay was used to detect expression levels of ribosomal protein S6 kinase (pS6k) and mTOR. To downregulate the expression of PTEN and cluster of differentiation 44 (CD44), a cell-surface glycoprotein, simultaneously, PTEN siRNA and CD44 siRNA sequences were mixed in equal proportions in co-interference experiments. RT-PCR detected the transcription level of CD44, and the relationship between the CD44 and axonal growth was observed after 48 h of interference. RESULTS: Microtubule-associated protein 2 (MAP2) expression was enhanced after three days of induction in SH-SY5Y cells. RT-PCR showed the transcription level of PTEN was significantly downregulated after 24 h of PTEN knockdown. mTOR and pS6k protein expression levels were significantly upregulated after 36 h of interference. CD44 transcription levels were upregulated after PTEN gene interference. The neurite length of the cells in the experimental interference group was significantly longer than that in the control group, and the expression level of CD44 was positively correlated with neurite growth. The neurite length of the PTEN-only interference group was significantly greater than that of the co-interference and ATRA groups. CONCLUSION: Activation of the mTOR pathway promoted neurite growth through upregulation of CD44 expression, thus promoting neuronal regeneration. |
format | Online Article Text |
id | pubmed-10288410 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | SAGE Publications |
record_format | MEDLINE/PubMed |
spelling | pubmed-102884102023-06-24 Activation of the mTOR pathway promotes neurite growth through upregulation of CD44 expression Zhang, Jiwei Gan, Wenjuan Peng, Ru Lu, Lei Lu, Weiqing Liu, Jiamei J Int Med Res Pre-Clinical Research Report OBJECTIVE: To explore the intrinsic mechanism of the mammalian target of rapamycin (mTOR) pathway activation and promotion of neuronal axon growth. METHODS: Human neuroblastoma cells, SH-SY5Y, were induced with all-trans retinoic acid (ATRA; 10 μM for three days) which differentiated the cell line into a neuronal-like state. Immunohistochemical staining was used to detect the differentiation status of the neuronal-like cells. Phosphatase and tensin homolog (PTEN) RNA interference (RNAi) experiments were performed on the differentiated cells; reverse transcription-polymerase chain reaction (RT-PCR) detected transcriptional levels of PTEN following 24 h of interference. After 36 h, western blot assay was used to detect expression levels of ribosomal protein S6 kinase (pS6k) and mTOR. To downregulate the expression of PTEN and cluster of differentiation 44 (CD44), a cell-surface glycoprotein, simultaneously, PTEN siRNA and CD44 siRNA sequences were mixed in equal proportions in co-interference experiments. RT-PCR detected the transcription level of CD44, and the relationship between the CD44 and axonal growth was observed after 48 h of interference. RESULTS: Microtubule-associated protein 2 (MAP2) expression was enhanced after three days of induction in SH-SY5Y cells. RT-PCR showed the transcription level of PTEN was significantly downregulated after 24 h of PTEN knockdown. mTOR and pS6k protein expression levels were significantly upregulated after 36 h of interference. CD44 transcription levels were upregulated after PTEN gene interference. The neurite length of the cells in the experimental interference group was significantly longer than that in the control group, and the expression level of CD44 was positively correlated with neurite growth. The neurite length of the PTEN-only interference group was significantly greater than that of the co-interference and ATRA groups. CONCLUSION: Activation of the mTOR pathway promoted neurite growth through upregulation of CD44 expression, thus promoting neuronal regeneration. SAGE Publications 2023-06-20 /pmc/articles/PMC10288410/ /pubmed/37340717 http://dx.doi.org/10.1177/03000605231178510 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by-nc/4.0/Creative Commons Non Commercial CC BY-NC: This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage). |
spellingShingle | Pre-Clinical Research Report Zhang, Jiwei Gan, Wenjuan Peng, Ru Lu, Lei Lu, Weiqing Liu, Jiamei Activation of the mTOR pathway promotes neurite growth through upregulation of CD44 expression |
title | Activation of the mTOR pathway promotes neurite growth through upregulation of CD44 expression |
title_full | Activation of the mTOR pathway promotes neurite growth through upregulation of CD44 expression |
title_fullStr | Activation of the mTOR pathway promotes neurite growth through upregulation of CD44 expression |
title_full_unstemmed | Activation of the mTOR pathway promotes neurite growth through upregulation of CD44 expression |
title_short | Activation of the mTOR pathway promotes neurite growth through upregulation of CD44 expression |
title_sort | activation of the mtor pathway promotes neurite growth through upregulation of cd44 expression |
topic | Pre-Clinical Research Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10288410/ https://www.ncbi.nlm.nih.gov/pubmed/37340717 http://dx.doi.org/10.1177/03000605231178510 |
work_keys_str_mv | AT zhangjiwei activationofthemtorpathwaypromotesneuritegrowththroughupregulationofcd44expression AT ganwenjuan activationofthemtorpathwaypromotesneuritegrowththroughupregulationofcd44expression AT pengru activationofthemtorpathwaypromotesneuritegrowththroughupregulationofcd44expression AT lulei activationofthemtorpathwaypromotesneuritegrowththroughupregulationofcd44expression AT luweiqing activationofthemtorpathwaypromotesneuritegrowththroughupregulationofcd44expression AT liujiamei activationofthemtorpathwaypromotesneuritegrowththroughupregulationofcd44expression |