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Downregulation of LPXN expression by siRNA decreases the malignant proliferation and transmembrane invasion of SHI-1 cells

The aim of the present study was to investigate the effects of decreasing leupaxin (LPXN) expression on the proliferation and invasion of human acute monocytic leukemia SHI-1 cells. The transfection efficiency of fluorescein amidite (FAM)-small interfering RNA (siRNA) was determined using flow cytom...

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Autores principales: Zhu, Guo-Hua, Dai, Hai-Ping, Shen, Qun, Zhang, Qi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6313184/
https://www.ncbi.nlm.nih.gov/pubmed/30655748
http://dx.doi.org/10.3892/ol.2018.9605
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author Zhu, Guo-Hua
Dai, Hai-Ping
Shen, Qun
Zhang, Qi
author_facet Zhu, Guo-Hua
Dai, Hai-Ping
Shen, Qun
Zhang, Qi
author_sort Zhu, Guo-Hua
collection PubMed
description The aim of the present study was to investigate the effects of decreasing leupaxin (LPXN) expression on the proliferation and invasion of human acute monocytic leukemia SHI-1 cells. The transfection efficiency of fluorescein amidite (FAM)-small interfering RNA (siRNA) was determined using flow cytometry, and the protein expression levels of LPXN, phosphorylated (p)-c-Jun N-terminal kinase (JNK), p-p38 mitogen-activated protein kinase (p38 MAPK) and p-extracellular-signal-regulated kinase (ERK) were detected by western blot analysis. Proliferation was determined using the cell counting kit-8 reagent and cellular transmembrane invasion ability was determined using a Transwell chamber system. The gelatinase levels of matrix metalloproteinase (MMP)-2 and MMP-9 in the cell culture supernatant were also analyzed by gelatin zymography. In SHI-1 cells, the optimal transfection conditions of siRNA were a cell density of 4×10(5) cells/ml and a ratio of siRNA/Lipofectamine(®) 2000 of 200 pmol/1 µl. The highest transfection efficiency of FAM-siRNA was 74.5%. In the present study, L2-siRNA was selected to effectively decrease the expression of LPXN. Following downregulation of LPXN expression by L2-siRNA, proliferation inhibition rates increased to 27.043±2.051 and cell transmembrane invasion rates decreased to 25.270±2.145 (P<0.05). The results of the western blot analysis and the gelatin zymography indicated that downregulation of LPXN expression increased the expression of p-p38 MAPK and p-JNK, and attenuated the secretion levels of MMP-2 and MMP-9. However, downregulation of LPXN expression had no effect on p-ERK expression in SHI-1 cells. The results of the present study indicated that downregulation of LPXN expression decreased the malignant proliferation and transmembrane invasion of SHI-1 cells by activating JNK and p38 MAPK, and inhibiting MMP-2 and MMP-9 secretion.
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spelling pubmed-63131842019-01-17 Downregulation of LPXN expression by siRNA decreases the malignant proliferation and transmembrane invasion of SHI-1 cells Zhu, Guo-Hua Dai, Hai-Ping Shen, Qun Zhang, Qi Oncol Lett Articles The aim of the present study was to investigate the effects of decreasing leupaxin (LPXN) expression on the proliferation and invasion of human acute monocytic leukemia SHI-1 cells. The transfection efficiency of fluorescein amidite (FAM)-small interfering RNA (siRNA) was determined using flow cytometry, and the protein expression levels of LPXN, phosphorylated (p)-c-Jun N-terminal kinase (JNK), p-p38 mitogen-activated protein kinase (p38 MAPK) and p-extracellular-signal-regulated kinase (ERK) were detected by western blot analysis. Proliferation was determined using the cell counting kit-8 reagent and cellular transmembrane invasion ability was determined using a Transwell chamber system. The gelatinase levels of matrix metalloproteinase (MMP)-2 and MMP-9 in the cell culture supernatant were also analyzed by gelatin zymography. In SHI-1 cells, the optimal transfection conditions of siRNA were a cell density of 4×10(5) cells/ml and a ratio of siRNA/Lipofectamine(®) 2000 of 200 pmol/1 µl. The highest transfection efficiency of FAM-siRNA was 74.5%. In the present study, L2-siRNA was selected to effectively decrease the expression of LPXN. Following downregulation of LPXN expression by L2-siRNA, proliferation inhibition rates increased to 27.043±2.051 and cell transmembrane invasion rates decreased to 25.270±2.145 (P<0.05). The results of the western blot analysis and the gelatin zymography indicated that downregulation of LPXN expression increased the expression of p-p38 MAPK and p-JNK, and attenuated the secretion levels of MMP-2 and MMP-9. However, downregulation of LPXN expression had no effect on p-ERK expression in SHI-1 cells. The results of the present study indicated that downregulation of LPXN expression decreased the malignant proliferation and transmembrane invasion of SHI-1 cells by activating JNK and p38 MAPK, and inhibiting MMP-2 and MMP-9 secretion. D.A. Spandidos 2019-01 2018-10-22 /pmc/articles/PMC6313184/ /pubmed/30655748 http://dx.doi.org/10.3892/ol.2018.9605 Text en Copyright: © Zhu et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
spellingShingle Articles
Zhu, Guo-Hua
Dai, Hai-Ping
Shen, Qun
Zhang, Qi
Downregulation of LPXN expression by siRNA decreases the malignant proliferation and transmembrane invasion of SHI-1 cells
title Downregulation of LPXN expression by siRNA decreases the malignant proliferation and transmembrane invasion of SHI-1 cells
title_full Downregulation of LPXN expression by siRNA decreases the malignant proliferation and transmembrane invasion of SHI-1 cells
title_fullStr Downregulation of LPXN expression by siRNA decreases the malignant proliferation and transmembrane invasion of SHI-1 cells
title_full_unstemmed Downregulation of LPXN expression by siRNA decreases the malignant proliferation and transmembrane invasion of SHI-1 cells
title_short Downregulation of LPXN expression by siRNA decreases the malignant proliferation and transmembrane invasion of SHI-1 cells
title_sort downregulation of lpxn expression by sirna decreases the malignant proliferation and transmembrane invasion of shi-1 cells
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6313184/
https://www.ncbi.nlm.nih.gov/pubmed/30655748
http://dx.doi.org/10.3892/ol.2018.9605
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