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Lncap-AI prostate cancer cell line establishment by Flutamide and androgen-free environment to promote cell adherent

BACKGROUND: To establish castration-resistant prostate cancer (CRPC) - Lncap androgen-independent (AI) cell line from Lncap androgen-dependent (AD) cell line, and explore the different molecular biological between these two cell lines. METHODS: The Lncap-AD cell line was cultured and passaged 60 tim...

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Autores principales: Wang, Huifeng, Wei, Xihua, Zhang, Die, Li, Weidong, Hu, Yanling
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9706963/
https://www.ncbi.nlm.nih.gov/pubmed/36443669
http://dx.doi.org/10.1186/s12860-022-00453-2
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author Wang, Huifeng
Wei, Xihua
Zhang, Die
Li, Weidong
Hu, Yanling
author_facet Wang, Huifeng
Wei, Xihua
Zhang, Die
Li, Weidong
Hu, Yanling
author_sort Wang, Huifeng
collection PubMed
description BACKGROUND: To establish castration-resistant prostate cancer (CRPC) - Lncap androgen-independent (AI) cell line from Lncap androgen-dependent (AD) cell line, and explore the different molecular biological between these two cell lines. METHODS: The Lncap-AD cell line was cultured and passaged 60 times over 16 months. The morphology of the Lncap-AI cell line was observed. AR levels identification were detected in qRT-PCR and Western Blot assay. CCK-8, EdU assay, wound healing assay and cell adhesion assays were used to observe the ability of proliferation, migration, and adhesion. SEM and TEM were used to observe microculture structure. At last, the PSA secrete ability was evaluated by Elisa assay. RESULTS: The Lncap-AD cell line was cultured and passaged 60 times over 16 months. The Lncap-AI cell line showed a morphologic change at the end stage of culture, the cells turned slender and cell space turned separated compared to the Lncap-AD cell line. The relative levels of AR-related genes in the Lncap-AI cell line were up-regulation compared to the Lncap-AD cell line both in mRNA and protein levels. The expression of AR and HK2 proteins were influenced and down-regulation by Enzalutamide in the Lncap-AD cell line, but no obvious difference in Lncap-AI cell lines. Lncap-AI cell line showed strong viability of proliferation, migration, and adhesion by CCK-8, EdU assay, wound healing assay, and adhesion assay. The microstructure of Scanning Electron Microscopy (SEM) showed many synapses in the Lncap-AI cell line and PC3 cell line, but not in the Lncap-AD cell line. At last, the PSA secrete ability was evaluated by Elisa assay, and PCa cell lines showed no significant difference. CONCLUSION: Simulation of CRPC progression, Lncap-AD cell line turned to Lncap-AI cell line with androgen deprivation therapy. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12860-022-00453-2.
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spelling pubmed-97069632022-11-30 Lncap-AI prostate cancer cell line establishment by Flutamide and androgen-free environment to promote cell adherent Wang, Huifeng Wei, Xihua Zhang, Die Li, Weidong Hu, Yanling BMC Mol Cell Biol Research BACKGROUND: To establish castration-resistant prostate cancer (CRPC) - Lncap androgen-independent (AI) cell line from Lncap androgen-dependent (AD) cell line, and explore the different molecular biological between these two cell lines. METHODS: The Lncap-AD cell line was cultured and passaged 60 times over 16 months. The morphology of the Lncap-AI cell line was observed. AR levels identification were detected in qRT-PCR and Western Blot assay. CCK-8, EdU assay, wound healing assay and cell adhesion assays were used to observe the ability of proliferation, migration, and adhesion. SEM and TEM were used to observe microculture structure. At last, the PSA secrete ability was evaluated by Elisa assay. RESULTS: The Lncap-AD cell line was cultured and passaged 60 times over 16 months. The Lncap-AI cell line showed a morphologic change at the end stage of culture, the cells turned slender and cell space turned separated compared to the Lncap-AD cell line. The relative levels of AR-related genes in the Lncap-AI cell line were up-regulation compared to the Lncap-AD cell line both in mRNA and protein levels. The expression of AR and HK2 proteins were influenced and down-regulation by Enzalutamide in the Lncap-AD cell line, but no obvious difference in Lncap-AI cell lines. Lncap-AI cell line showed strong viability of proliferation, migration, and adhesion by CCK-8, EdU assay, wound healing assay, and adhesion assay. The microstructure of Scanning Electron Microscopy (SEM) showed many synapses in the Lncap-AI cell line and PC3 cell line, but not in the Lncap-AD cell line. At last, the PSA secrete ability was evaluated by Elisa assay, and PCa cell lines showed no significant difference. CONCLUSION: Simulation of CRPC progression, Lncap-AD cell line turned to Lncap-AI cell line with androgen deprivation therapy. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12860-022-00453-2. BioMed Central 2022-11-28 /pmc/articles/PMC9706963/ /pubmed/36443669 http://dx.doi.org/10.1186/s12860-022-00453-2 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Wang, Huifeng
Wei, Xihua
Zhang, Die
Li, Weidong
Hu, Yanling
Lncap-AI prostate cancer cell line establishment by Flutamide and androgen-free environment to promote cell adherent
title Lncap-AI prostate cancer cell line establishment by Flutamide and androgen-free environment to promote cell adherent
title_full Lncap-AI prostate cancer cell line establishment by Flutamide and androgen-free environment to promote cell adherent
title_fullStr Lncap-AI prostate cancer cell line establishment by Flutamide and androgen-free environment to promote cell adherent
title_full_unstemmed Lncap-AI prostate cancer cell line establishment by Flutamide and androgen-free environment to promote cell adherent
title_short Lncap-AI prostate cancer cell line establishment by Flutamide and androgen-free environment to promote cell adherent
title_sort lncap-ai prostate cancer cell line establishment by flutamide and androgen-free environment to promote cell adherent
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9706963/
https://www.ncbi.nlm.nih.gov/pubmed/36443669
http://dx.doi.org/10.1186/s12860-022-00453-2
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