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Increased cytomegalovirus replication by 5-Azacytidine and viral-induced cytoplasmic expression of DNMT‑1 in medulloblastoma and endothelial cells

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Estekizadeh, Atosa ; Landázur, Natalia ; Bartek, Jiri ; Beltoft Brøchner, Christian ; Davoudi, Belghis ; Broholm, Helle ; Karimi, Mohsen ; Ekström, Tomas J ; Rahbar, Afsar. / Increased cytomegalovirus replication by 5-Azacytidine and viral-induced cytoplasmic expression of DNMT‑1 in medulloblastoma and endothelial cells. I: International Journal of Oncology. 2018 ; Bind 52, Nr. 4. s. 1317-1327.

Bibtex

@article{d63ef54a62bb410280783f1fbf1e065f,
title = "Increased cytomegalovirus replication by 5-Azacytidine and viral-induced cytoplasmic expression of DNMT‑1 in medulloblastoma and endothelial cells",
abstract = "Among all brain tumors diagnosed in children, medulloblastomas (MBs) are associated with a poor prognosis. The etiology of MB is not fully understood, yet the impact of epigenetic alterations of oncogenes has previously been established. During the past decade, the human cytomegalovirus (HCMV) has been detected in several types of cancer, including MB. Since DNA methylation occurs in the cell nucleus and this is considered a host defence response, we studied the impact of HCMV infection on DNA methyltransferase (DNMT‑1) in MB (D324) cells, human umbilical vein endothelial cells (HUVECs) as well as in MB tissue sections. We hypothesized that infection and DNMT‑1 intracellular localization are linked. Uninfected and HCMV‑infected D324 cells and HUVECs were analyzed for HCMV immediate early (HCMV‑IE) protein, HCMV‑glycoprotein B (HCMV‑gB) and DNMT‑1 using immunofluorescence staining and quantitative ELISA. DNMT‑1 localized to the nucleus of uninfected and HCMV‑IE- expressing D324 cells and HUVECs, but accumulated in the extra nuclear space in all HCMV‑gB-positive cells. Inhibition of HCMV late protein expression by Cymevene{\textregistered} (ganciclovir) prevented the cytoplasmic localization of DNMT‑1. Treatment of HCMV‑ infected D324 cells and HUVECs with the methylation inhibitor 5-Azacytidine (5AZA), significantly increased HCMV‑IE and HCMV‑gB gene transcription and protein expression. Immunohistochemical staining of DNMT‑1 and HCMV proteins in MB cancer tissue sections revealed both nuclear and cytoplasmic DNMT‑1 localization. In conclusion, DNMT‑1 resides in the cytoplasm of HCMV‑gB-expressing HUVECs and D324 cells. Increased viral protein synthesis in 5AZA-treated cells suggests that HCMV replication may benefit from a DNA methyltransferase-free cellular environment. Our findings emphasize the importance of assessing potential viral activation in the treatment of MB patients with epigenetic drugs.",
keywords = "Antimetabolites, Antineoplastic/pharmacology, Azacitidine/pharmacology, Cerebellar Neoplasms/virology, Cytomegalovirus, Cytomegalovirus Infections/complications, Cytoplasm/metabolism, DNA (Cytosine-5-)-Methyltransferase 1/biosynthesis, Human Umbilical Vein Endothelial Cells/virology, Humans, Medulloblastoma/virology, Viral Proteins, Virus Activation/drug effects, Virus Replication/drug effects",
author = "Atosa Estekizadeh and Natalia Land{\'a}zur and Jiri Bartek and {Beltoft Br{\o}chner}, Christian and Belghis Davoudi and Helle Broholm and Mohsen Karimi and Ekstr{\"o}m, {Tomas J} and Afsar Rahbar",
year = "2018",
month = apr,
doi = "10.3892/ijo.2018.4286",
language = "English",
volume = "52",
pages = "1317--1327",
journal = "International Journal of Oncology",
issn = "1019-6439",
publisher = "Demetrios A./Spandidos Ed. & Pub",
number = "4",

}

RIS

TY - JOUR

T1 - Increased cytomegalovirus replication by 5-Azacytidine and viral-induced cytoplasmic expression of DNMT‑1 in medulloblastoma and endothelial cells

AU - Estekizadeh, Atosa

AU - Landázur, Natalia

AU - Bartek, Jiri

AU - Beltoft Brøchner, Christian

AU - Davoudi, Belghis

AU - Broholm, Helle

AU - Karimi, Mohsen

AU - Ekström, Tomas J

AU - Rahbar, Afsar

PY - 2018/4

Y1 - 2018/4

N2 - Among all brain tumors diagnosed in children, medulloblastomas (MBs) are associated with a poor prognosis. The etiology of MB is not fully understood, yet the impact of epigenetic alterations of oncogenes has previously been established. During the past decade, the human cytomegalovirus (HCMV) has been detected in several types of cancer, including MB. Since DNA methylation occurs in the cell nucleus and this is considered a host defence response, we studied the impact of HCMV infection on DNA methyltransferase (DNMT‑1) in MB (D324) cells, human umbilical vein endothelial cells (HUVECs) as well as in MB tissue sections. We hypothesized that infection and DNMT‑1 intracellular localization are linked. Uninfected and HCMV‑infected D324 cells and HUVECs were analyzed for HCMV immediate early (HCMV‑IE) protein, HCMV‑glycoprotein B (HCMV‑gB) and DNMT‑1 using immunofluorescence staining and quantitative ELISA. DNMT‑1 localized to the nucleus of uninfected and HCMV‑IE- expressing D324 cells and HUVECs, but accumulated in the extra nuclear space in all HCMV‑gB-positive cells. Inhibition of HCMV late protein expression by Cymevene® (ganciclovir) prevented the cytoplasmic localization of DNMT‑1. Treatment of HCMV‑ infected D324 cells and HUVECs with the methylation inhibitor 5-Azacytidine (5AZA), significantly increased HCMV‑IE and HCMV‑gB gene transcription and protein expression. Immunohistochemical staining of DNMT‑1 and HCMV proteins in MB cancer tissue sections revealed both nuclear and cytoplasmic DNMT‑1 localization. In conclusion, DNMT‑1 resides in the cytoplasm of HCMV‑gB-expressing HUVECs and D324 cells. Increased viral protein synthesis in 5AZA-treated cells suggests that HCMV replication may benefit from a DNA methyltransferase-free cellular environment. Our findings emphasize the importance of assessing potential viral activation in the treatment of MB patients with epigenetic drugs.

AB - Among all brain tumors diagnosed in children, medulloblastomas (MBs) are associated with a poor prognosis. The etiology of MB is not fully understood, yet the impact of epigenetic alterations of oncogenes has previously been established. During the past decade, the human cytomegalovirus (HCMV) has been detected in several types of cancer, including MB. Since DNA methylation occurs in the cell nucleus and this is considered a host defence response, we studied the impact of HCMV infection on DNA methyltransferase (DNMT‑1) in MB (D324) cells, human umbilical vein endothelial cells (HUVECs) as well as in MB tissue sections. We hypothesized that infection and DNMT‑1 intracellular localization are linked. Uninfected and HCMV‑infected D324 cells and HUVECs were analyzed for HCMV immediate early (HCMV‑IE) protein, HCMV‑glycoprotein B (HCMV‑gB) and DNMT‑1 using immunofluorescence staining and quantitative ELISA. DNMT‑1 localized to the nucleus of uninfected and HCMV‑IE- expressing D324 cells and HUVECs, but accumulated in the extra nuclear space in all HCMV‑gB-positive cells. Inhibition of HCMV late protein expression by Cymevene® (ganciclovir) prevented the cytoplasmic localization of DNMT‑1. Treatment of HCMV‑ infected D324 cells and HUVECs with the methylation inhibitor 5-Azacytidine (5AZA), significantly increased HCMV‑IE and HCMV‑gB gene transcription and protein expression. Immunohistochemical staining of DNMT‑1 and HCMV proteins in MB cancer tissue sections revealed both nuclear and cytoplasmic DNMT‑1 localization. In conclusion, DNMT‑1 resides in the cytoplasm of HCMV‑gB-expressing HUVECs and D324 cells. Increased viral protein synthesis in 5AZA-treated cells suggests that HCMV replication may benefit from a DNA methyltransferase-free cellular environment. Our findings emphasize the importance of assessing potential viral activation in the treatment of MB patients with epigenetic drugs.

KW - Antimetabolites, Antineoplastic/pharmacology

KW - Azacitidine/pharmacology

KW - Cerebellar Neoplasms/virology

KW - Cytomegalovirus

KW - Cytomegalovirus Infections/complications

KW - Cytoplasm/metabolism

KW - DNA (Cytosine-5-)-Methyltransferase 1/biosynthesis

KW - Human Umbilical Vein Endothelial Cells/virology

KW - Humans

KW - Medulloblastoma/virology

KW - Viral Proteins

KW - Virus Activation/drug effects

KW - Virus Replication/drug effects

U2 - 10.3892/ijo.2018.4286

DO - 10.3892/ijo.2018.4286

M3 - Journal article

C2 - 29484388

VL - 52

SP - 1317

EP - 1327

JO - International Journal of Oncology

JF - International Journal of Oncology

SN - 1019-6439

IS - 4

ER -

ID: 55351128