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PML silencing inhibits cell proliferation and induces DNA damage in cultured ovarian cancer cells

The promyelocytic leukemia (PML) gene is a tumor suppressor gene. It was first identified in acute promyelocytic leukemia, in which it is fused to retinoic acid receptor α by the (15;17) chromosomal translocation. The function of the PML protein is frequently lost or aberrant in human solid tumors....

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Autores principales: Liu, Sheng-Bing, Shen, Zhong-Fei, Guo, Yan-Jun, Cao, Li-Xian, Xu, Ying
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5492820/
https://www.ncbi.nlm.nih.gov/pubmed/28685056
http://dx.doi.org/10.3892/br.2017.919
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author Liu, Sheng-Bing
Shen, Zhong-Fei
Guo, Yan-Jun
Cao, Li-Xian
Xu, Ying
author_facet Liu, Sheng-Bing
Shen, Zhong-Fei
Guo, Yan-Jun
Cao, Li-Xian
Xu, Ying
author_sort Liu, Sheng-Bing
collection PubMed
description The promyelocytic leukemia (PML) gene is a tumor suppressor gene. It was first identified in acute promyelocytic leukemia, in which it is fused to retinoic acid receptor α by the (15;17) chromosomal translocation. The function of the PML protein is frequently lost or aberrant in human solid tumors. In human ovarian carcinoma tissue, PML detected by immunohistochemistry was highly expressed. A PML-silencing vector, pSRG-shPml, was constructed and used to transfect human ovarian cancer cells. Cells were cultured and selected with puromycin for 10–15 days, and then the PML mRNA expression levels were detected by RT-qPCR and immunofluorescence. Proliferation and clone number of PML-depleted cells were detected using MTT assay and colony-forming assay. The protein expression associated with DNA damage and apoptosis was assessed in PML-depleted cells using western blot analysis and immunofluorescence. The results showed that PML was highly expressed in human ovarian tissue. The proliferation and colony formation of ovarian cancer cells were significantly inhibited after PML was depleted. Western blot analysis and immunofluorescence revealed that p-H2AX and cleaved caspase-3 expression significantly increased after PML silencing. PML was located in the nucleus, and it formed foci after X-ray irradiation. PML foci increased significantly with increasing irradiation doses.
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spelling pubmed-54928202017-07-06 PML silencing inhibits cell proliferation and induces DNA damage in cultured ovarian cancer cells Liu, Sheng-Bing Shen, Zhong-Fei Guo, Yan-Jun Cao, Li-Xian Xu, Ying Biomed Rep Articles The promyelocytic leukemia (PML) gene is a tumor suppressor gene. It was first identified in acute promyelocytic leukemia, in which it is fused to retinoic acid receptor α by the (15;17) chromosomal translocation. The function of the PML protein is frequently lost or aberrant in human solid tumors. In human ovarian carcinoma tissue, PML detected by immunohistochemistry was highly expressed. A PML-silencing vector, pSRG-shPml, was constructed and used to transfect human ovarian cancer cells. Cells were cultured and selected with puromycin for 10–15 days, and then the PML mRNA expression levels were detected by RT-qPCR and immunofluorescence. Proliferation and clone number of PML-depleted cells were detected using MTT assay and colony-forming assay. The protein expression associated with DNA damage and apoptosis was assessed in PML-depleted cells using western blot analysis and immunofluorescence. The results showed that PML was highly expressed in human ovarian tissue. The proliferation and colony formation of ovarian cancer cells were significantly inhibited after PML was depleted. Western blot analysis and immunofluorescence revealed that p-H2AX and cleaved caspase-3 expression significantly increased after PML silencing. PML was located in the nucleus, and it formed foci after X-ray irradiation. PML foci increased significantly with increasing irradiation doses. D.A. Spandidos 2017-07 2017-05-24 /pmc/articles/PMC5492820/ /pubmed/28685056 http://dx.doi.org/10.3892/br.2017.919 Text en Copyright: © Liu 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
Liu, Sheng-Bing
Shen, Zhong-Fei
Guo, Yan-Jun
Cao, Li-Xian
Xu, Ying
PML silencing inhibits cell proliferation and induces DNA damage in cultured ovarian cancer cells
title PML silencing inhibits cell proliferation and induces DNA damage in cultured ovarian cancer cells
title_full PML silencing inhibits cell proliferation and induces DNA damage in cultured ovarian cancer cells
title_fullStr PML silencing inhibits cell proliferation and induces DNA damage in cultured ovarian cancer cells
title_full_unstemmed PML silencing inhibits cell proliferation and induces DNA damage in cultured ovarian cancer cells
title_short PML silencing inhibits cell proliferation and induces DNA damage in cultured ovarian cancer cells
title_sort pml silencing inhibits cell proliferation and induces dna damage in cultured ovarian cancer cells
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5492820/
https://www.ncbi.nlm.nih.gov/pubmed/28685056
http://dx.doi.org/10.3892/br.2017.919
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