Detection of human papillomavirus genotypes, herpes simplex, varicella zoster and cytomegalovirus in breast cancer patients

Abstract

Background: The role of viruses as a cause of breast cancer (BC) has been significantly investigated in recent years. Human papillomavirus (HPV) has been detected in invasive breast carcinomas, while most studies have only focused on the detection of viral DNA, we aimed to examine the prevalence and genotypes of HPV among Iranian BC patients. We also examined the presence of herpes simplex-1 (HSV-1), herpes simplex-2 (HSV-2), varicella zoster virus (VZV), and cytomegalovirus (CMV) in these samples.

Methods: We collected and analyzed 70 Formalin-Fixed Paraffin-Embedded (FFPE) blocks including 59 BC samples, and 11 benign breast lesions as control from Iranian patients using nested PCR. Real-time PCR utilized as a confirming test to nested PCR findings. Genotyping of HPV positive samples was performed, the samples were also subjected to a multiplex PCR to detect HSV-1, HSV-2, VZV, and CMV in BC.

Results: Papillomavirus DNA was present in 7 of 59 BC samples (11.8%); while none was detected in control samples. The most prevalent type was HPV18, followed by HPV 6. All HPV positive patients had high tumor grades (II/ III) with a histologic diagnosis of ductal carcinoma. The patient age range was 33 to 73 years with a median of 51 years. Most of HPV positive patients had low levels of education. HPV16 was not detected. Also, 5 of 59 BC specimens (8.47%), were positive for HSV-1. But none of the samples were positive for HSV-2, VZV, and CMV.

Conclusions: Our results suggest a carcinogenesis role for High-risk HPV (HPV18) in breast tumors. Our findings of HSV-1 and low-risk HPV (HPV6) in BCs may propose a cancer-causing role for them. Further large-scale studies are warranted to assess the significance of our findings.

Keywords: Breast cancer; Genotype; HPV; Herpes virus; Human papillomavirus; PCR.

Fig. 1

An agarose gel electrophoretogram of amplified GAPDH after the PCR as described in the materials and methods. Lanes 19 to 28 breast cancer samples, last lane: 100-bp DNA ladder

Fig. 2

An agarose gel electrophoretogram of amplified DNA after the nested PCR as described in the materials and methods. C+ Lane: positive control (hpv18 HeLa cell line), Lanes 1 to 13 breast cancer samples, C− lane: negative control, last lane: 100-bp DNA ladder

Fig. 3

Gel electrophoresis of the multiplex PCR amplification products. Lanes 19 to 26 breast cancer samples. The amplicon length was 269 bp. C+ lane: positive control, C− lane: negative control, last lane: 100-bp DNA ladder

Fig. 4

Phylogenetic trees of HPV16 and HPV18 Based on L1 Nucleotide were Constructed Using neighbor joining method and the Kimura 2-Parameter model by MEGA 6 package