Cathepsin B Regulates Mice Granulosa Cells’ Apoptosis and Proliferation In Vitro

Cathepsin B (CTSB), a lysosomal cysteine protease’s high expression and activity, has been reported to cause poor-quality embryos in porcine and bovine. Nevertheless, CTSB functions in mice granulosa cells remain to explore. To discuss the CTSB...

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Autores principales:	Chao Chen, Muhammad Jamil Ahmad, Tingzhu Ye, Chao Du, Xinxin Zhang, Aixin Liang, Liguo Yang
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>CTSB</i> mice proliferation apoptosis granulosa cells Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/775c843a68584cab9d83a37449b12bcf
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spelling	oai:doaj.org-article:775c843a68584cab9d83a37449b12bcf2021-11-11T17:15:48ZCathepsin B Regulates Mice Granulosa Cells’ Apoptosis and Proliferation In Vitro10.3390/ijms2221118271422-00671661-6596https://doaj.org/article/775c843a68584cab9d83a37449b12bcf2021-10-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11827https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Cathepsin B (<i>CTSB</i>), a lysosomal cysteine protease’s high expression and activity, has been reported to cause poor-quality embryos in porcine and bovine. Nevertheless, <i>CTSB</i> functions in mice granulosa cells remain to explore. To discuss the <i>CTSB</i> functional role in follicular dynamics, we studied apoptosis, proliferation, cell cycle progression, and related signaling pathways in primary mouse granulosa cells transfected with small interference RNA specific to <i>CTSB</i> (siCTSB) for 48 h. Further, mRNA and protein expression of cell proliferation regulators (<i>Myc</i> and <i>cyclin D2</i>), apoptosis regulators (<i>caspase 3</i>, <i>caspase 8</i>, <i>TNF-α</i>, and <i>Bcl2</i>), steroidogenesis-related genes (<i>FSHR</i> and <i>CYP11A1</i>), and autophagy markers (<i>LC3-I</i> and <i>ATG5</i>) were investigated. In addition, the effect of <i>CTSB</i> on steroidogenesis and autophagy was also examined. Flow cytometry analysis assay displayed that silencing of <i>CTSB</i> decreased the early and total apoptosis rate by downregulating <i>TNF-α</i>, <i>caspase 8</i>, and <i>caspase 3</i>, and upregulating <i>Bcl2</i>. By regulating <i>Myc</i> and <i>cyclin D2</i> expression and activating the p-Akt and p-ERK pathways, <i>CTSB</i> knockdown increased GC proliferation and number. A significant decline in estradiol and progesterone concentrations was observed parallel to a significant decrease in autophagy-related markers <i>LC3-I</i> and <i>ATG5</i> compared to the control group. Herein, we demonstrated that <i>CTSB</i> serves as a proapoptotic agent and plays a critical role in folliculogenesis in female mice by mediating apoptosis, autophagy, proliferation, and steroidogenesis. Hence, <i>CTSB</i> could be a potential prognostic agent for female infertility.Chao ChenMuhammad Jamil AhmadTingzhu YeChao DuXinxin ZhangAixin LiangLiguo YangMDPI AGarticle<i>CTSB</i>miceproliferationapoptosisgranulosa cellsBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11827, p 11827 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>CTSB</i> mice proliferation apoptosis granulosa cells Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	<i>CTSB</i> mice proliferation apoptosis granulosa cells Biology (General) QH301-705.5 Chemistry QD1-999 Chao Chen Muhammad Jamil Ahmad Tingzhu Ye Chao Du Xinxin Zhang Aixin Liang Liguo Yang Cathepsin B Regulates Mice Granulosa Cells’ Apoptosis and Proliferation In Vitro
description	Cathepsin B (<i>CTSB</i>), a lysosomal cysteine protease’s high expression and activity, has been reported to cause poor-quality embryos in porcine and bovine. Nevertheless, <i>CTSB</i> functions in mice granulosa cells remain to explore. To discuss the <i>CTSB</i> functional role in follicular dynamics, we studied apoptosis, proliferation, cell cycle progression, and related signaling pathways in primary mouse granulosa cells transfected with small interference RNA specific to <i>CTSB</i> (siCTSB) for 48 h. Further, mRNA and protein expression of cell proliferation regulators (<i>Myc</i> and <i>cyclin D2</i>), apoptosis regulators (<i>caspase 3</i>, <i>caspase 8</i>, <i>TNF-α</i>, and <i>Bcl2</i>), steroidogenesis-related genes (<i>FSHR</i> and <i>CYP11A1</i>), and autophagy markers (<i>LC3-I</i> and <i>ATG5</i>) were investigated. In addition, the effect of <i>CTSB</i> on steroidogenesis and autophagy was also examined. Flow cytometry analysis assay displayed that silencing of <i>CTSB</i> decreased the early and total apoptosis rate by downregulating <i>TNF-α</i>, <i>caspase 8</i>, and <i>caspase 3</i>, and upregulating <i>Bcl2</i>. By regulating <i>Myc</i> and <i>cyclin D2</i> expression and activating the p-Akt and p-ERK pathways, <i>CTSB</i> knockdown increased GC proliferation and number. A significant decline in estradiol and progesterone concentrations was observed parallel to a significant decrease in autophagy-related markers <i>LC3-I</i> and <i>ATG5</i> compared to the control group. Herein, we demonstrated that <i>CTSB</i> serves as a proapoptotic agent and plays a critical role in folliculogenesis in female mice by mediating apoptosis, autophagy, proliferation, and steroidogenesis. Hence, <i>CTSB</i> could be a potential prognostic agent for female infertility.
format	article
author	Chao Chen Muhammad Jamil Ahmad Tingzhu Ye Chao Du Xinxin Zhang Aixin Liang Liguo Yang
author_facet	Chao Chen Muhammad Jamil Ahmad Tingzhu Ye Chao Du Xinxin Zhang Aixin Liang Liguo Yang
author_sort	Chao Chen
title	Cathepsin B Regulates Mice Granulosa Cells’ Apoptosis and Proliferation In Vitro
title_short	Cathepsin B Regulates Mice Granulosa Cells’ Apoptosis and Proliferation In Vitro
title_full	Cathepsin B Regulates Mice Granulosa Cells’ Apoptosis and Proliferation In Vitro
title_fullStr	Cathepsin B Regulates Mice Granulosa Cells’ Apoptosis and Proliferation In Vitro
title_full_unstemmed	Cathepsin B Regulates Mice Granulosa Cells’ Apoptosis and Proliferation In Vitro
title_sort	cathepsin b regulates mice granulosa cells’ apoptosis and proliferation in vitro
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/775c843a68584cab9d83a37449b12bcf
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_version_	1718432125372334080

Cathepsin B Regulates Mice Granulosa Cells’ Apoptosis and Proliferation In Vitro

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