CDX2/mir-145-5p/SENP1 Pathways Affect LNCaP Cells Invasion and Migration
Background/Aims: Recently, rapidly accumulating evidence has shown that microRNAs (miRNAs) are involved in human tumorigenesis, and the dysregulation of miRNAs has been observed in many cancers, including prostate cancer. miR-145-5p, an miRNA with reduced expression in prostate cancer cells, has bee...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2019-06-01
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| Series: | Frontiers in Oncology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fonc.2019.00477/full |
| _version_ | 1818989669969297408 |
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| author | Jin-Hua He Ze-Ping Han Mao-Xian Zou Meng-Ling He Yu-Guang Li Lei Zheng |
| author_facet | Jin-Hua He Ze-Ping Han Mao-Xian Zou Meng-Ling He Yu-Guang Li Lei Zheng |
| author_sort | Jin-Hua He |
| collection | DOAJ |
| description | Background/Aims: Recently, rapidly accumulating evidence has shown that microRNAs (miRNAs) are involved in human tumorigenesis, and the dysregulation of miRNAs has been observed in many cancers, including prostate cancer. miR-145-5p, an miRNA with reduced expression in prostate cancer cells, has been shown to have a tumor suppressive role in a variety of tumors. However, its underlying mechanism requires further elucidation.Methods: A lentiviral expression vector for miR-145-5p was constructed and used to establish a stable cell line (LNCaP) expressing miR-145-5p. The cells were cultured normally and divided into the control group (control), negative control group (negative control), and test group (miR-145-5p). Inhibition of proliferation was measured by a WST-8 assay. The early apoptosis rate of cells was detected by flow cytometry. Clone formation ability was detected by a clone formation inhibition test. Cell invasion and migration capacity was detected by a Transwell assay. The relative expression levels of proteins were detected by western blotting. We constructed a nude mouse model of prostate cancer to observe the effect of miR-145-5p on the growth of transplanted tumors. TargetScan bioinformatics software was used to predict target genes regulated by miR-14-5p. ChIPBase was used to predict transcription factors with binding sites in the upstream promoter region of miR-145-5p. Quantitative reverse transcription PCR was used to detect the relative expression level of genes. A bifluorescence-reporter gene vector was constructed to confirm the regulation of target genes by miR-145-5p. We used 5′ rapid amplification of cDNA ends to confirm the transcription start site of miR-145-5p.Chromatin immunoprecipitation technology was used to detect the effect of transcription factors binding to miR-145-5p.Results: The overexpression of miR-145-5p not only inhibited the proliferation, invasion, and migration of LNCaP cells but also promoted their early apoptosis. After overexpressing miR-145-5p, the expression of small ubiquitin-like modifier protein-specific protease 1 (SENP1), and caudal-related homeobox 2 (CDX2) protein was decreased in LNCaP cells. The transcription factor CDX2 bound to the miR-145-5p promoter region and inhibited its transcription. The transcription start site of miR-145-5p was located at a guanine residue 1,408 bp upstream of the stem-loop sequence. Upon overexpression, miR-145-5p could bind to the 3′-untranslated region of SENP1 to inhibit its translation.Conclusion: These results suggested that CDX2 inhibits the expression of miR-145-5p, thereby relieving the inhibitory effect of miR-145-5p on the translation of SENP1 and affecting the invasion and migration of prostate cancer cells. |
| first_indexed | 2024-12-20T19:42:10Z |
| format | Article |
| id | doaj.art-3d2af88ee2d14e93ae3c8fa11cb24b98 |
| institution | Directory Open Access Journal |
| issn | 2234-943X |
| language | English |
| last_indexed | 2024-12-20T19:42:10Z |
| publishDate | 2019-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Oncology |
| spelling | doaj.art-3d2af88ee2d14e93ae3c8fa11cb24b982022-12-21T19:28:29ZengFrontiers Media S.A.Frontiers in Oncology2234-943X2019-06-01910.3389/fonc.2019.00477457639CDX2/mir-145-5p/SENP1 Pathways Affect LNCaP Cells Invasion and MigrationJin-Hua He0Ze-Ping Han1Mao-Xian Zou2Meng-Ling He3Yu-Guang Li4Lei Zheng5Department of Laboratory Medicine, Central Hospital of Panyu District, Guangzhou, ChinaDepartment of Laboratory Medicine, Central Hospital of Panyu District, Guangzhou, ChinaDepartment of Laboratory Medicine, Central Hospital of Panyu District, Guangzhou, ChinaDepartment of Laboratory Medicine, Central Hospital of Panyu District, Guangzhou, ChinaDepartment of Laboratory Medicine, Central Hospital of Panyu District, Guangzhou, ChinaDepartment of Laboratory Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, ChinaBackground/Aims: Recently, rapidly accumulating evidence has shown that microRNAs (miRNAs) are involved in human tumorigenesis, and the dysregulation of miRNAs has been observed in many cancers, including prostate cancer. miR-145-5p, an miRNA with reduced expression in prostate cancer cells, has been shown to have a tumor suppressive role in a variety of tumors. However, its underlying mechanism requires further elucidation.Methods: A lentiviral expression vector for miR-145-5p was constructed and used to establish a stable cell line (LNCaP) expressing miR-145-5p. The cells were cultured normally and divided into the control group (control), negative control group (negative control), and test group (miR-145-5p). Inhibition of proliferation was measured by a WST-8 assay. The early apoptosis rate of cells was detected by flow cytometry. Clone formation ability was detected by a clone formation inhibition test. Cell invasion and migration capacity was detected by a Transwell assay. The relative expression levels of proteins were detected by western blotting. We constructed a nude mouse model of prostate cancer to observe the effect of miR-145-5p on the growth of transplanted tumors. TargetScan bioinformatics software was used to predict target genes regulated by miR-14-5p. ChIPBase was used to predict transcription factors with binding sites in the upstream promoter region of miR-145-5p. Quantitative reverse transcription PCR was used to detect the relative expression level of genes. A bifluorescence-reporter gene vector was constructed to confirm the regulation of target genes by miR-145-5p. We used 5′ rapid amplification of cDNA ends to confirm the transcription start site of miR-145-5p.Chromatin immunoprecipitation technology was used to detect the effect of transcription factors binding to miR-145-5p.Results: The overexpression of miR-145-5p not only inhibited the proliferation, invasion, and migration of LNCaP cells but also promoted their early apoptosis. After overexpressing miR-145-5p, the expression of small ubiquitin-like modifier protein-specific protease 1 (SENP1), and caudal-related homeobox 2 (CDX2) protein was decreased in LNCaP cells. The transcription factor CDX2 bound to the miR-145-5p promoter region and inhibited its transcription. The transcription start site of miR-145-5p was located at a guanine residue 1,408 bp upstream of the stem-loop sequence. Upon overexpression, miR-145-5p could bind to the 3′-untranslated region of SENP1 to inhibit its translation.Conclusion: These results suggested that CDX2 inhibits the expression of miR-145-5p, thereby relieving the inhibitory effect of miR-145-5p on the translation of SENP1 and affecting the invasion and migration of prostate cancer cells.https://www.frontiersin.org/article/10.3389/fonc.2019.00477/fullmiR-145-5pCDX2SENP1prostate cancerinvasionmigration |
| spellingShingle | Jin-Hua He Ze-Ping Han Mao-Xian Zou Meng-Ling He Yu-Guang Li Lei Zheng CDX2/mir-145-5p/SENP1 Pathways Affect LNCaP Cells Invasion and Migration Frontiers in Oncology miR-145-5p CDX2 SENP1 prostate cancer invasion migration |
| title | CDX2/mir-145-5p/SENP1 Pathways Affect LNCaP Cells Invasion and Migration |
| title_full | CDX2/mir-145-5p/SENP1 Pathways Affect LNCaP Cells Invasion and Migration |
| title_fullStr | CDX2/mir-145-5p/SENP1 Pathways Affect LNCaP Cells Invasion and Migration |
| title_full_unstemmed | CDX2/mir-145-5p/SENP1 Pathways Affect LNCaP Cells Invasion and Migration |
| title_short | CDX2/mir-145-5p/SENP1 Pathways Affect LNCaP Cells Invasion and Migration |
| title_sort | cdx2 mir 145 5p senp1 pathways affect lncap cells invasion and migration |
| topic | miR-145-5p CDX2 SENP1 prostate cancer invasion migration |
| url | https://www.frontiersin.org/article/10.3389/fonc.2019.00477/full |
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