CRISPR/cas9-mediated overexpression of long non-coding RNA SRY-box transcription factor 21 antisense divergent transcript 1 regulates the proliferation of osteosarcoma by increasing the expression of mechanistic target of rapamycin kinase and Kruppel like factor 4

Osteosarcoma is derived from primitive mesenchymal cells and is the most common primary solid malignant tumor of bone. The cause of osteosarcoma is unclear. In recent years, the role of non-coding sequences in regulating protein expression in tumors has been paid more and more attention, especially...

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Autores principales:	Weiying Zhang, Qiang Wang, Haibo Du, Shichao Jiang
Formato:	article
Lenguaje:	EN
Publicado:	Taylor & Francis Group 2021
Materias:	osteosarcoma lncrna sox21-as1 mtor klf4 hsa-mir-7-5p hsa-mir-145-5p Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/cee7dfa879014c659edae445c2f9e415
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spelling	oai:doaj.org-article:cee7dfa879014c659edae445c2f9e4152021-11-04T15:51:54ZCRISPR/cas9-mediated overexpression of long non-coding RNA SRY-box transcription factor 21 antisense divergent transcript 1 regulates the proliferation of osteosarcoma by increasing the expression of mechanistic target of rapamycin kinase and Kruppel like factor 42165-59792165-598710.1080/21655979.2021.1995106https://doaj.org/article/cee7dfa879014c659edae445c2f9e4152021-10-01T00:00:00Zhttp://dx.doi.org/10.1080/21655979.2021.1995106https://doaj.org/toc/2165-5979https://doaj.org/toc/2165-5987Osteosarcoma is derived from primitive mesenchymal cells and is the most common primary solid malignant tumor of bone. The cause of osteosarcoma is unclear. In recent years, the role of non-coding sequences in regulating protein expression in tumors has been paid more and more attention, especially long non-coding RNA (lncRNA). We speculate that SRY-box transcription factor 21 antisense divergent transcript 1 (SOX21-AS1) can regulate the expression of the mechanistic target of rapamycin kinase (mTOR) and Kruppel like factor 4 (KLF4) through sponge hsa-mir-7-5p and hsa-mir-145-5p. We knocked lncRNA SOX21-AS1 into the genome of 143B through CRISPR/Cas9, then screened out a monoclonal cell line. Detect the transcription level and protein expression level of the above-mentioned related genes, and cell proliferation. Then, ginsenoside Rg3 was added to culture the cell line knocked into lncRNA SOX21-AS1, and the expression levels of lncRNA SOX21-AS1, hsa-mir-7-5p, miR-145-5P, mTOR and KLF4 were detected by RT-qPCR and Western blot. Cell proliferation method detects cell viability, explores the molecular mechanism of lncRNA SOX21-AS1 in osteosarcoma, and whether it can be used as a potential drug target for the treatment of osteosarcoma. Our results demonstrate that the overexpression of lncRNA SOX21-AS1 up-regulates mTOR and KLF4 by sponging hsa-mir-7-5p and hsa-mir-145-5p, and ultimately regulates the proliferation of osteosarcoma. And proved that ginsenoside Rg3 can inhibit the cell proliferation of osteosarcoma by reducing the expression level of lncRNA SOX21-AS1. It provides an alternative for the treatment of osteosarcoma in the future.Weiying ZhangQiang WangHaibo DuShichao JiangTaylor & Francis Grouparticleosteosarcomalncrna sox21-as1mtorklf4hsa-mir-7-5phsa-mir-145-5pBiotechnologyTP248.13-248.65ENBioengineered, Vol 0, Iss 0 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	osteosarcoma lncrna sox21-as1 mtor klf4 hsa-mir-7-5p hsa-mir-145-5p Biotechnology TP248.13-248.65
spellingShingle	osteosarcoma lncrna sox21-as1 mtor klf4 hsa-mir-7-5p hsa-mir-145-5p Biotechnology TP248.13-248.65 Weiying Zhang Qiang Wang Haibo Du Shichao Jiang CRISPR/cas9-mediated overexpression of long non-coding RNA SRY-box transcription factor 21 antisense divergent transcript 1 regulates the proliferation of osteosarcoma by increasing the expression of mechanistic target of rapamycin kinase and Kruppel like factor 4
description	Osteosarcoma is derived from primitive mesenchymal cells and is the most common primary solid malignant tumor of bone. The cause of osteosarcoma is unclear. In recent years, the role of non-coding sequences in regulating protein expression in tumors has been paid more and more attention, especially long non-coding RNA (lncRNA). We speculate that SRY-box transcription factor 21 antisense divergent transcript 1 (SOX21-AS1) can regulate the expression of the mechanistic target of rapamycin kinase (mTOR) and Kruppel like factor 4 (KLF4) through sponge hsa-mir-7-5p and hsa-mir-145-5p. We knocked lncRNA SOX21-AS1 into the genome of 143B through CRISPR/Cas9, then screened out a monoclonal cell line. Detect the transcription level and protein expression level of the above-mentioned related genes, and cell proliferation. Then, ginsenoside Rg3 was added to culture the cell line knocked into lncRNA SOX21-AS1, and the expression levels of lncRNA SOX21-AS1, hsa-mir-7-5p, miR-145-5P, mTOR and KLF4 were detected by RT-qPCR and Western blot. Cell proliferation method detects cell viability, explores the molecular mechanism of lncRNA SOX21-AS1 in osteosarcoma, and whether it can be used as a potential drug target for the treatment of osteosarcoma. Our results demonstrate that the overexpression of lncRNA SOX21-AS1 up-regulates mTOR and KLF4 by sponging hsa-mir-7-5p and hsa-mir-145-5p, and ultimately regulates the proliferation of osteosarcoma. And proved that ginsenoside Rg3 can inhibit the cell proliferation of osteosarcoma by reducing the expression level of lncRNA SOX21-AS1. It provides an alternative for the treatment of osteosarcoma in the future.
format	article
author	Weiying Zhang Qiang Wang Haibo Du Shichao Jiang
author_facet	Weiying Zhang Qiang Wang Haibo Du Shichao Jiang
author_sort	Weiying Zhang
title	CRISPR/cas9-mediated overexpression of long non-coding RNA SRY-box transcription factor 21 antisense divergent transcript 1 regulates the proliferation of osteosarcoma by increasing the expression of mechanistic target of rapamycin kinase and Kruppel like factor 4
title_short	CRISPR/cas9-mediated overexpression of long non-coding RNA SRY-box transcription factor 21 antisense divergent transcript 1 regulates the proliferation of osteosarcoma by increasing the expression of mechanistic target of rapamycin kinase and Kruppel like factor 4
title_full	CRISPR/cas9-mediated overexpression of long non-coding RNA SRY-box transcription factor 21 antisense divergent transcript 1 regulates the proliferation of osteosarcoma by increasing the expression of mechanistic target of rapamycin kinase and Kruppel like factor 4
title_fullStr	CRISPR/cas9-mediated overexpression of long non-coding RNA SRY-box transcription factor 21 antisense divergent transcript 1 regulates the proliferation of osteosarcoma by increasing the expression of mechanistic target of rapamycin kinase and Kruppel like factor 4
title_full_unstemmed	CRISPR/cas9-mediated overexpression of long non-coding RNA SRY-box transcription factor 21 antisense divergent transcript 1 regulates the proliferation of osteosarcoma by increasing the expression of mechanistic target of rapamycin kinase and Kruppel like factor 4
title_sort	crispr/cas9-mediated overexpression of long non-coding rna sry-box transcription factor 21 antisense divergent transcript 1 regulates the proliferation of osteosarcoma by increasing the expression of mechanistic target of rapamycin kinase and kruppel like factor 4
publisher	Taylor & Francis Group
publishDate	2021
url	https://doaj.org/article/cee7dfa879014c659edae445c2f9e415
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CRISPR/cas9-mediated overexpression of long non-coding RNA SRY-box transcription factor 21 antisense divergent transcript 1 regulates the proliferation of osteosarcoma by increasing the expression of mechanistic target of rapamycin kinase and Kruppel like factor 4

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