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Human papillomavirus and Epstein-Barr virus co-infection in cervical carcinoma in Algerian women.

Khenchouche A, Sadouki N, Boudriche A, Houali K, Graba A, Ooka T, Bouguermouh A - Virol. J. (2013)

Bottom Line: The highest co-infection (HR-HPV and EBV) was found in squamous cell carcinoma cases (67%).The latter cases showed 27% and 29% expression of EBV BARF-1 and LMP-1 oncogenes respectively.This could be taken into account as bad prognosis in this type of cancer.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratoire de Virologie Moléculaire, UMR5537, C,N,R,S, Faculté de Médecine R,T,H, Laennec, Lyon, France. tadamasa.ooka@univ-lyon1.fr.

ABSTRACT

Background: Despite the fact that the implication of human papillomavirus (HPV) in the carcinogenesis and prognosis of cervical cancer is well established, the impact of a co-infection with high risk HPV (HR-HPV) and Epstein-Barr virus (EBV) is still not fully understood.

Methods: Fifty eight randomly selected cases of squamous cell carcinomas (SCC) of the uterine cervix, 14 normal cervices specimens, 21 CIN-2/3 and 16 CIN-1 cases were examined for EBV and HPV infections. Detection of HR-HPV specific sequences was carried out by PCR amplification using consensus primers of Manos and by Digene Hybrid Capture. The presence of EBV was revealed by amplifying a 660 bp specific EBV sequence of BALF1. mRNA expression of LMP-1 in one hand and protein levels of BARF-1, LMP-1 and EBNA-1 in the other hand were assessed by RT-PCR and immunoblotting and/or immunohischemistry respectively.

Results: HR-HPV infection was found in patients with SCC (88%), low-grade (75%) and high grade (95%) lesions compared to only 14% of normal cervix cases. However, 69%, 12.5%, 38.1%, and 14% of SCC, CIN-1, CIN-2/3 and normal cervix tissues, respectively, were EBV infected. The highest co-infection (HR-HPV and EBV) was found in squamous cell carcinoma cases (67%). The latter cases showed 27% and 29% expression of EBV BARF-1 and LMP-1 oncogenes respectively.

Conclusion: The high rate of HR-HPV and EBV co-infection in SCC suggests that EBV infection is incriminated in cervical cancer progression. This could be taken into account as bad prognosis in this type of cancer. However, the mode of action in dual infection in cervical oncogenesis needs further investigation.

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Detection of BARF-1 protein by Western blotting. Immunoblotting of BARF-1 on seven cervical tumor biopsies. Fifty micrograms of protein extract as measured by a Biorad protein assay were deposited onto 12% polyacrylamide gel and electrophoresed. Proteins were transferred onto nitrocellulose paper. BARF1 protein encoding 29 kDa (p29) was revealed by PepIII rabbit polyclonal anti-BARF-1. BARF-1 protein (p29) produced by the BARF-1 recombinant adenovirus system was used as a positive control indicated in first lane as BARF1.
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Figure 4: Detection of BARF-1 protein by Western blotting. Immunoblotting of BARF-1 on seven cervical tumor biopsies. Fifty micrograms of protein extract as measured by a Biorad protein assay were deposited onto 12% polyacrylamide gel and electrophoresed. Proteins were transferred onto nitrocellulose paper. BARF1 protein encoding 29 kDa (p29) was revealed by PepIII rabbit polyclonal anti-BARF-1. BARF-1 protein (p29) produced by the BARF-1 recombinant adenovirus system was used as a positive control indicated in first lane as BARF1.

Mentions: As shown in Figure 3, the LMP-1 transcript was about 200 bp which was spliced from the 406 bp DNA sequence (see Material and Methods). Using immunoblotting technique, BARF-1 protein was detected in only 21.7% (5/23) of SCC biopsies analyzed from 50 μg cellular extract (Figure 4). We also examined the transcriptional and translational expression of HPV and EBV oncogenes (LMP-1 and BARF-1) by immunohistochemistry (IHC) (Figure 5). IHC analysis has allowed the identification of HPV and EBV products. As shown in Figure 5D, HPV was expressed in SCC biopsies. EBV antigens, EBNA-1 and LMP-1, were present as a brown nuclear and membrane staining (Figure 5B and C respectively). EBNA-1 protein was detected in 34.7% (8/23). While IHC analysis showed 26% (6/23) LMP-1 positive cases (Figure 5C). Negative squamous carcinoma cells for EBNA-1 and LMP-1 proteins are shown in Figures 5E and F respectively.


Human papillomavirus and Epstein-Barr virus co-infection in cervical carcinoma in Algerian women.

Khenchouche A, Sadouki N, Boudriche A, Houali K, Graba A, Ooka T, Bouguermouh A - Virol. J. (2013)

Detection of BARF-1 protein by Western blotting. Immunoblotting of BARF-1 on seven cervical tumor biopsies. Fifty micrograms of protein extract as measured by a Biorad protein assay were deposited onto 12% polyacrylamide gel and electrophoresed. Proteins were transferred onto nitrocellulose paper. BARF1 protein encoding 29 kDa (p29) was revealed by PepIII rabbit polyclonal anti-BARF-1. BARF-1 protein (p29) produced by the BARF-1 recombinant adenovirus system was used as a positive control indicated in first lane as BARF1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4225508&req=5

Figure 4: Detection of BARF-1 protein by Western blotting. Immunoblotting of BARF-1 on seven cervical tumor biopsies. Fifty micrograms of protein extract as measured by a Biorad protein assay were deposited onto 12% polyacrylamide gel and electrophoresed. Proteins were transferred onto nitrocellulose paper. BARF1 protein encoding 29 kDa (p29) was revealed by PepIII rabbit polyclonal anti-BARF-1. BARF-1 protein (p29) produced by the BARF-1 recombinant adenovirus system was used as a positive control indicated in first lane as BARF1.
Mentions: As shown in Figure 3, the LMP-1 transcript was about 200 bp which was spliced from the 406 bp DNA sequence (see Material and Methods). Using immunoblotting technique, BARF-1 protein was detected in only 21.7% (5/23) of SCC biopsies analyzed from 50 μg cellular extract (Figure 4). We also examined the transcriptional and translational expression of HPV and EBV oncogenes (LMP-1 and BARF-1) by immunohistochemistry (IHC) (Figure 5). IHC analysis has allowed the identification of HPV and EBV products. As shown in Figure 5D, HPV was expressed in SCC biopsies. EBV antigens, EBNA-1 and LMP-1, were present as a brown nuclear and membrane staining (Figure 5B and C respectively). EBNA-1 protein was detected in 34.7% (8/23). While IHC analysis showed 26% (6/23) LMP-1 positive cases (Figure 5C). Negative squamous carcinoma cells for EBNA-1 and LMP-1 proteins are shown in Figures 5E and F respectively.

Bottom Line: The highest co-infection (HR-HPV and EBV) was found in squamous cell carcinoma cases (67%).The latter cases showed 27% and 29% expression of EBV BARF-1 and LMP-1 oncogenes respectively.This could be taken into account as bad prognosis in this type of cancer.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratoire de Virologie Moléculaire, UMR5537, C,N,R,S, Faculté de Médecine R,T,H, Laennec, Lyon, France. tadamasa.ooka@univ-lyon1.fr.

ABSTRACT

Background: Despite the fact that the implication of human papillomavirus (HPV) in the carcinogenesis and prognosis of cervical cancer is well established, the impact of a co-infection with high risk HPV (HR-HPV) and Epstein-Barr virus (EBV) is still not fully understood.

Methods: Fifty eight randomly selected cases of squamous cell carcinomas (SCC) of the uterine cervix, 14 normal cervices specimens, 21 CIN-2/3 and 16 CIN-1 cases were examined for EBV and HPV infections. Detection of HR-HPV specific sequences was carried out by PCR amplification using consensus primers of Manos and by Digene Hybrid Capture. The presence of EBV was revealed by amplifying a 660 bp specific EBV sequence of BALF1. mRNA expression of LMP-1 in one hand and protein levels of BARF-1, LMP-1 and EBNA-1 in the other hand were assessed by RT-PCR and immunoblotting and/or immunohischemistry respectively.

Results: HR-HPV infection was found in patients with SCC (88%), low-grade (75%) and high grade (95%) lesions compared to only 14% of normal cervix cases. However, 69%, 12.5%, 38.1%, and 14% of SCC, CIN-1, CIN-2/3 and normal cervix tissues, respectively, were EBV infected. The highest co-infection (HR-HPV and EBV) was found in squamous cell carcinoma cases (67%). The latter cases showed 27% and 29% expression of EBV BARF-1 and LMP-1 oncogenes respectively.

Conclusion: The high rate of HR-HPV and EBV co-infection in SCC suggests that EBV infection is incriminated in cervical cancer progression. This could be taken into account as bad prognosis in this type of cancer. However, the mode of action in dual infection in cervical oncogenesis needs further investigation.

Show MeSH
Related in: MedlinePlus