Induction of Chromosomal Instability in Colonic Cells by the Human Polyomavirus JC Virus

Luigi Ricciardiello, Michele Baglioni, Catia Giovannini, Milena Pariali, Giovanna Cenacchi, Alessandro Ripalti, Maria Paola Landini, Hirofumi Sawa, Kazuo Nagashima, Richard J. Frisque, Ajay Goel, C. Richard Boland, Mauro Tognon, Enrico Roda, Franco Bazzoli

Research output: Contribution to journalArticle

93 Citations (Scopus)

Abstract

Most colorectal cancers display chromosomal instability, which is characterized by gross chromosomal rearrangements, loss of heterozygosity and aneuploidy. We have previously demonstrated a link between JC virus strains Mad-1 and Δ98 and colorectal cancer. Others have also associated the virus to the induction of colon cancer and aneuploid brain tumors by producing a highly tumorigenic protein named T antigen (TAg), which binds to β-catenin and inactivates key proteins such as p53. The aim is to demonstrate that JC virus is capable of inducing chromosomal instability in colonic cells. We used the human colon cancer cell line RKO as a model. The cell line has wild-type p53, wild-type β-catenin and APC and is diploid. Neuroblastoma JCI cells, which are infected with the virus, VA13 fibroblasts, which are transformed by the SV40 TAg, were used as positive controls. HCT116, which has mutated β-catenin, and SW480, which is a model of CIN, were also used as controls. The genomes of the Mad-1 and Δ98 strains were transfected into cells. As negative controls we used pUC or no plasmids. Cells were collected at 0, 7, 14, and 21 days after transfection. PCR was used for the detection of TAg and the regulatory region DNA sequences at different time frames and Southern blot of whole genomic extracts for viral DNA integration into the host genome. Immunofluorescence and Western blot were performed for TAg, viral capsid proteins, and nuclear β-catenin expressions, whereas coimmunoprecipitation was used to detect protein interactions. Karyotype analysis and electron microscopy were performed to seek chromosomal instability and cell abnormalities, respectively. Retention of viral sequences was observed for Mad-1- and Δ98-transfected RKO cells at all time frames with PCR only, whereas Southern blot analysis showed nonintegrated sequences at T7 alone. TAg and capsid protein expressions, as well as increased p53 and nuclear β-catenin, were observed between T0 and T7 for Mad-1 and Δ98 alone. Also, interaction between TAg and both p53 and β-catenin was also observed between T0 and T7. Chromosomal instability, characterized by chromosomal breakage, dicentric chromosomes, and increasing ploidy, was observed at all time frames for Mad-1 and Δ98, as well as cell abnormalities. In conclusion, we demonstrate that JC virus Mad-1 and Δ98 are able to induce chromosomal instability in colonic cells with a hit and run mechanism that involves an early interaction with β-catenin and p53.

Original languageEnglish (US)
Pages (from-to)7256-7262
Number of pages7
JournalCancer Research
Volume63
Issue number21
StatePublished - Nov 1 2003

Fingerprint

JC Virus
Chromosomal Instability
Catenins
Viral Tumor Antigens
Capsid Proteins
Aneuploidy
Southern Blotting
Brain Neoplasms
Colonic Neoplasms
Colorectal Neoplasms
Genome
Virus Integration
Polyomavirus Transforming Antigens
Satellite Viruses
Virus Activation
Chromosome Breakage
Cell Line
Polymerase Chain Reaction
Proteins
Ploidies

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

Ricciardiello, L., Baglioni, M., Giovannini, C., Pariali, M., Cenacchi, G., Ripalti, A., ... Bazzoli, F. (2003). Induction of Chromosomal Instability in Colonic Cells by the Human Polyomavirus JC Virus. Cancer Research, 63(21), 7256-7262.
Ricciardiello, Luigi ; Baglioni, Michele ; Giovannini, Catia ; Pariali, Milena ; Cenacchi, Giovanna ; Ripalti, Alessandro ; Landini, Maria Paola ; Sawa, Hirofumi ; Nagashima, Kazuo ; Frisque, Richard J. ; Goel, Ajay ; Boland, C. Richard ; Tognon, Mauro ; Roda, Enrico ; Bazzoli, Franco. / Induction of Chromosomal Instability in Colonic Cells by the Human Polyomavirus JC Virus. In: Cancer Research. 2003 ; Vol. 63, No. 21. pp. 7256-7262.
@article{080b0f6d50d3412f8ae8a4f2fbd56f6c,
title = "Induction of Chromosomal Instability in Colonic Cells by the Human Polyomavirus JC Virus",
abstract = "Most colorectal cancers display chromosomal instability, which is characterized by gross chromosomal rearrangements, loss of heterozygosity and aneuploidy. We have previously demonstrated a link between JC virus strains Mad-1 and Δ98 and colorectal cancer. Others have also associated the virus to the induction of colon cancer and aneuploid brain tumors by producing a highly tumorigenic protein named T antigen (TAg), which binds to β-catenin and inactivates key proteins such as p53. The aim is to demonstrate that JC virus is capable of inducing chromosomal instability in colonic cells. We used the human colon cancer cell line RKO as a model. The cell line has wild-type p53, wild-type β-catenin and APC and is diploid. Neuroblastoma JCI cells, which are infected with the virus, VA13 fibroblasts, which are transformed by the SV40 TAg, were used as positive controls. HCT116, which has mutated β-catenin, and SW480, which is a model of CIN, were also used as controls. The genomes of the Mad-1 and Δ98 strains were transfected into cells. As negative controls we used pUC or no plasmids. Cells were collected at 0, 7, 14, and 21 days after transfection. PCR was used for the detection of TAg and the regulatory region DNA sequences at different time frames and Southern blot of whole genomic extracts for viral DNA integration into the host genome. Immunofluorescence and Western blot were performed for TAg, viral capsid proteins, and nuclear β-catenin expressions, whereas coimmunoprecipitation was used to detect protein interactions. Karyotype analysis and electron microscopy were performed to seek chromosomal instability and cell abnormalities, respectively. Retention of viral sequences was observed for Mad-1- and Δ98-transfected RKO cells at all time frames with PCR only, whereas Southern blot analysis showed nonintegrated sequences at T7 alone. TAg and capsid protein expressions, as well as increased p53 and nuclear β-catenin, were observed between T0 and T7 for Mad-1 and Δ98 alone. Also, interaction between TAg and both p53 and β-catenin was also observed between T0 and T7. Chromosomal instability, characterized by chromosomal breakage, dicentric chromosomes, and increasing ploidy, was observed at all time frames for Mad-1 and Δ98, as well as cell abnormalities. In conclusion, we demonstrate that JC virus Mad-1 and Δ98 are able to induce chromosomal instability in colonic cells with a hit and run mechanism that involves an early interaction with β-catenin and p53.",
author = "Luigi Ricciardiello and Michele Baglioni and Catia Giovannini and Milena Pariali and Giovanna Cenacchi and Alessandro Ripalti and Landini, {Maria Paola} and Hirofumi Sawa and Kazuo Nagashima and Frisque, {Richard J.} and Ajay Goel and Boland, {C. Richard} and Mauro Tognon and Enrico Roda and Franco Bazzoli",
year = "2003",
month = "11",
day = "1",
language = "English (US)",
volume = "63",
pages = "7256--7262",
journal = "Journal of Cancer Research",
issn = "0008-5472",
publisher = "American Association for Cancer Research Inc.",
number = "21",

}

Ricciardiello, L, Baglioni, M, Giovannini, C, Pariali, M, Cenacchi, G, Ripalti, A, Landini, MP, Sawa, H, Nagashima, K, Frisque, RJ, Goel, A, Boland, CR, Tognon, M, Roda, E & Bazzoli, F 2003, 'Induction of Chromosomal Instability in Colonic Cells by the Human Polyomavirus JC Virus', Cancer Research, vol. 63, no. 21, pp. 7256-7262.

Induction of Chromosomal Instability in Colonic Cells by the Human Polyomavirus JC Virus. / Ricciardiello, Luigi; Baglioni, Michele; Giovannini, Catia; Pariali, Milena; Cenacchi, Giovanna; Ripalti, Alessandro; Landini, Maria Paola; Sawa, Hirofumi; Nagashima, Kazuo; Frisque, Richard J.; Goel, Ajay; Boland, C. Richard; Tognon, Mauro; Roda, Enrico; Bazzoli, Franco.

In: Cancer Research, Vol. 63, No. 21, 01.11.2003, p. 7256-7262.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Induction of Chromosomal Instability in Colonic Cells by the Human Polyomavirus JC Virus

AU - Ricciardiello, Luigi

AU - Baglioni, Michele

AU - Giovannini, Catia

AU - Pariali, Milena

AU - Cenacchi, Giovanna

AU - Ripalti, Alessandro

AU - Landini, Maria Paola

AU - Sawa, Hirofumi

AU - Nagashima, Kazuo

AU - Frisque, Richard J.

AU - Goel, Ajay

AU - Boland, C. Richard

AU - Tognon, Mauro

AU - Roda, Enrico

AU - Bazzoli, Franco

PY - 2003/11/1

Y1 - 2003/11/1

N2 - Most colorectal cancers display chromosomal instability, which is characterized by gross chromosomal rearrangements, loss of heterozygosity and aneuploidy. We have previously demonstrated a link between JC virus strains Mad-1 and Δ98 and colorectal cancer. Others have also associated the virus to the induction of colon cancer and aneuploid brain tumors by producing a highly tumorigenic protein named T antigen (TAg), which binds to β-catenin and inactivates key proteins such as p53. The aim is to demonstrate that JC virus is capable of inducing chromosomal instability in colonic cells. We used the human colon cancer cell line RKO as a model. The cell line has wild-type p53, wild-type β-catenin and APC and is diploid. Neuroblastoma JCI cells, which are infected with the virus, VA13 fibroblasts, which are transformed by the SV40 TAg, were used as positive controls. HCT116, which has mutated β-catenin, and SW480, which is a model of CIN, were also used as controls. The genomes of the Mad-1 and Δ98 strains were transfected into cells. As negative controls we used pUC or no plasmids. Cells were collected at 0, 7, 14, and 21 days after transfection. PCR was used for the detection of TAg and the regulatory region DNA sequences at different time frames and Southern blot of whole genomic extracts for viral DNA integration into the host genome. Immunofluorescence and Western blot were performed for TAg, viral capsid proteins, and nuclear β-catenin expressions, whereas coimmunoprecipitation was used to detect protein interactions. Karyotype analysis and electron microscopy were performed to seek chromosomal instability and cell abnormalities, respectively. Retention of viral sequences was observed for Mad-1- and Δ98-transfected RKO cells at all time frames with PCR only, whereas Southern blot analysis showed nonintegrated sequences at T7 alone. TAg and capsid protein expressions, as well as increased p53 and nuclear β-catenin, were observed between T0 and T7 for Mad-1 and Δ98 alone. Also, interaction between TAg and both p53 and β-catenin was also observed between T0 and T7. Chromosomal instability, characterized by chromosomal breakage, dicentric chromosomes, and increasing ploidy, was observed at all time frames for Mad-1 and Δ98, as well as cell abnormalities. In conclusion, we demonstrate that JC virus Mad-1 and Δ98 are able to induce chromosomal instability in colonic cells with a hit and run mechanism that involves an early interaction with β-catenin and p53.

AB - Most colorectal cancers display chromosomal instability, which is characterized by gross chromosomal rearrangements, loss of heterozygosity and aneuploidy. We have previously demonstrated a link between JC virus strains Mad-1 and Δ98 and colorectal cancer. Others have also associated the virus to the induction of colon cancer and aneuploid brain tumors by producing a highly tumorigenic protein named T antigen (TAg), which binds to β-catenin and inactivates key proteins such as p53. The aim is to demonstrate that JC virus is capable of inducing chromosomal instability in colonic cells. We used the human colon cancer cell line RKO as a model. The cell line has wild-type p53, wild-type β-catenin and APC and is diploid. Neuroblastoma JCI cells, which are infected with the virus, VA13 fibroblasts, which are transformed by the SV40 TAg, were used as positive controls. HCT116, which has mutated β-catenin, and SW480, which is a model of CIN, were also used as controls. The genomes of the Mad-1 and Δ98 strains were transfected into cells. As negative controls we used pUC or no plasmids. Cells were collected at 0, 7, 14, and 21 days after transfection. PCR was used for the detection of TAg and the regulatory region DNA sequences at different time frames and Southern blot of whole genomic extracts for viral DNA integration into the host genome. Immunofluorescence and Western blot were performed for TAg, viral capsid proteins, and nuclear β-catenin expressions, whereas coimmunoprecipitation was used to detect protein interactions. Karyotype analysis and electron microscopy were performed to seek chromosomal instability and cell abnormalities, respectively. Retention of viral sequences was observed for Mad-1- and Δ98-transfected RKO cells at all time frames with PCR only, whereas Southern blot analysis showed nonintegrated sequences at T7 alone. TAg and capsid protein expressions, as well as increased p53 and nuclear β-catenin, were observed between T0 and T7 for Mad-1 and Δ98 alone. Also, interaction between TAg and both p53 and β-catenin was also observed between T0 and T7. Chromosomal instability, characterized by chromosomal breakage, dicentric chromosomes, and increasing ploidy, was observed at all time frames for Mad-1 and Δ98, as well as cell abnormalities. In conclusion, we demonstrate that JC virus Mad-1 and Δ98 are able to induce chromosomal instability in colonic cells with a hit and run mechanism that involves an early interaction with β-catenin and p53.

UR - http://www.scopus.com/inward/record.url?scp=10744227638&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=10744227638&partnerID=8YFLogxK

M3 - Article

VL - 63

SP - 7256

EP - 7262

JO - Journal of Cancer Research

JF - Journal of Cancer Research

SN - 0008-5472

IS - 21

ER -

Ricciardiello L, Baglioni M, Giovannini C, Pariali M, Cenacchi G, Ripalti A et al. Induction of Chromosomal Instability in Colonic Cells by the Human Polyomavirus JC Virus. Cancer Research. 2003 Nov 1;63(21):7256-7262.