Silencing of LINC00467 inhibits cell proliferation in testicular germ cell tumors cells

A significant decrease in LINC00467 expression in testicular germ cell tumors (TGCTs) was found in our previous study in comparison to adjacent tissue. Interestingly, the expression of LINC00467 correlated with the pathological grade of the tumor in TGCT patients. The higher the expression of LINC00...

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Main Authors: Fang Zhu, Zhizhong Liu, Qianyin Zhou, Dai Zhou, Jingyu Fan, Hao Bo, Liqing Fan
Format: Article
Language:English
Published: Association of Basic Medical Sciences of Federation of Bosnia and Herzegovina 2023-09-01
Series:Biomolecules & Biomedicine
Subjects:
Online Access:https://www.bjbms.org/ojs/index.php/bjbms/article/view/8969
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author Fang Zhu
Zhizhong Liu
Qianyin Zhou
Dai Zhou
Jingyu Fan
Hao Bo
Liqing Fan
author_facet Fang Zhu
Zhizhong Liu
Qianyin Zhou
Dai Zhou
Jingyu Fan
Hao Bo
Liqing Fan
author_sort Fang Zhu
collection DOAJ
description A significant decrease in LINC00467 expression in testicular germ cell tumors (TGCTs) was found in our previous study in comparison to adjacent tissue. Interestingly, the expression of LINC00467 correlated with the pathological grade of the tumor in TGCT patients. The higher the expression of LINC00467 was, the worse the prognosis of the patients with TGCT was. Despite these findings, the exact role of LINC00467 in the development of TGCTs requires further investigation. LINC00467 expression was downregulated in the NCCIT and TCam-2 cell lines via small interfering RNA (siRNA) silencing. The levels of gene expression were validated using quantitative real-time polymerase chain reaction (qRT-PCR) analyses. Cell proliferation was evaluated by the MTT and Cell Counting Kit-8 (CCK8) assays, whereas flow cytometry was used to assess the effects on the cell cycle. Western blotting analysis was used to detect expression levels of protein. Additionally, RNA-sequencing and bioinformatics methods were used to investigate the mechanism of action of LINC00467 in TGCTs. The suppression of LINC00467 expression resulted in decreased cell proliferation and induced S-phase arrest. Furthermore, the suppression of LINC00467 downregulated proliferating cell nuclear antigen (PCNA), a protein related to cell cycle regulation, while it upregulated p21 expression. In other studies involving dihydrotestosterone (DHT) stimulation, it was observed that DHT could upregulate LINC00467 expression. In addition, silencing of the LINC00467 reversed the effect of testosterone on cell proliferation. The Gene Set Enrichment Analysis (GSEA) revealed that LINC00467 regulated the p53 pathway by modulating the expression of CCNG1. Our study found that LINC00467 regulates cell proliferation by inducing S-phase arrest through the cell cycle-related proteins PCNA and p21. These findings contribute to our understanding of non-coding RNAs mechanisms involved in the development of TGCTs.
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spelling doaj.art-f44d3316e70b42d285b83bc96bb8977d2024-03-15T13:22:28ZengAssociation of Basic Medical Sciences of Federation of Bosnia and HerzegovinaBiomolecules & Biomedicine2831-08962831-090X2023-09-0123510.17305/bb.2023.8969Silencing of LINC00467 inhibits cell proliferation in testicular germ cell tumors cellsFang Zhu0https://orcid.org/0000-0002-8289-7362Zhizhong Liu1Qianyin Zhou2Dai Zhou3Jingyu Fan4Hao Bo5Liqing Fan6https://orcid.org/0000-0002-8289-7362Institute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, ChinaInstitute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, China; Department of Urology, Hunan Cancer Hospital, The Affiliated Cancer Hospital of Xiangya, School of Medicine of Central South University, Changsha, Hunan, ChinaInstitute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, ChinaInstitute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, China; Clinical Research Center for Reproduction and Genetics in Hunan Province, Reproductive and Genetic Hospital of CITIC-Xiangya, Changsha, ChinaDepartment of Chemistry and Biochemistry, University of South Carolina, Columbia, SC, United StatesInstitute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, China; Clinical Research Center for Reproduction and Genetics in Hunan Province, Reproductive and Genetic Hospital of CITIC-Xiangya, Changsha, ChinaInstitute of Reproductive and Stem Cell Engineering, School of Basic Medical Science, Central South University, Changsha, China; Clinical Research Center for Reproduction and Genetics in Hunan Province, Reproductive and Genetic Hospital of CITIC-Xiangya, Changsha, ChinaA significant decrease in LINC00467 expression in testicular germ cell tumors (TGCTs) was found in our previous study in comparison to adjacent tissue. Interestingly, the expression of LINC00467 correlated with the pathological grade of the tumor in TGCT patients. The higher the expression of LINC00467 was, the worse the prognosis of the patients with TGCT was. Despite these findings, the exact role of LINC00467 in the development of TGCTs requires further investigation. LINC00467 expression was downregulated in the NCCIT and TCam-2 cell lines via small interfering RNA (siRNA) silencing. The levels of gene expression were validated using quantitative real-time polymerase chain reaction (qRT-PCR) analyses. Cell proliferation was evaluated by the MTT and Cell Counting Kit-8 (CCK8) assays, whereas flow cytometry was used to assess the effects on the cell cycle. Western blotting analysis was used to detect expression levels of protein. Additionally, RNA-sequencing and bioinformatics methods were used to investigate the mechanism of action of LINC00467 in TGCTs. The suppression of LINC00467 expression resulted in decreased cell proliferation and induced S-phase arrest. Furthermore, the suppression of LINC00467 downregulated proliferating cell nuclear antigen (PCNA), a protein related to cell cycle regulation, while it upregulated p21 expression. In other studies involving dihydrotestosterone (DHT) stimulation, it was observed that DHT could upregulate LINC00467 expression. In addition, silencing of the LINC00467 reversed the effect of testosterone on cell proliferation. The Gene Set Enrichment Analysis (GSEA) revealed that LINC00467 regulated the p53 pathway by modulating the expression of CCNG1. Our study found that LINC00467 regulates cell proliferation by inducing S-phase arrest through the cell cycle-related proteins PCNA and p21. These findings contribute to our understanding of non-coding RNAs mechanisms involved in the development of TGCTs. https://www.bjbms.org/ojs/index.php/bjbms/article/view/8969Testicular germ cell tumors (TGCT)LINC0046dihydrotestosterone (DHT)cell proliferationcell cycle
spellingShingle Fang Zhu
Zhizhong Liu
Qianyin Zhou
Dai Zhou
Jingyu Fan
Hao Bo
Liqing Fan
Silencing of LINC00467 inhibits cell proliferation in testicular germ cell tumors cells
Biomolecules & Biomedicine
Testicular germ cell tumors (TGCT)
LINC0046
dihydrotestosterone (DHT)
cell proliferation
cell cycle
title Silencing of LINC00467 inhibits cell proliferation in testicular germ cell tumors cells
title_full Silencing of LINC00467 inhibits cell proliferation in testicular germ cell tumors cells
title_fullStr Silencing of LINC00467 inhibits cell proliferation in testicular germ cell tumors cells
title_full_unstemmed Silencing of LINC00467 inhibits cell proliferation in testicular germ cell tumors cells
title_short Silencing of LINC00467 inhibits cell proliferation in testicular germ cell tumors cells
title_sort silencing of linc00467 inhibits cell proliferation in testicular germ cell tumors cells
topic Testicular germ cell tumors (TGCT)
LINC0046
dihydrotestosterone (DHT)
cell proliferation
cell cycle
url https://www.bjbms.org/ojs/index.php/bjbms/article/view/8969
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