Detection and analysis of human papillomavirus (HPV) DNA in breast cancer patients by an effective method of HPV capture

PLoS One. 2014 Mar 10;9(3):e90343. doi: 10.1371/journal.pone.0090343. eCollection 2014.

Abstract

Despite an increase in the number of molecular epidemiological studies conducted in recent years to evaluate the association between human papillomavirus (HPV) and the risk of breast carcinoma, these studies remain inconclusive. Here we aim to detect HPV DNA in various tissues from patients with breast carcinoma using the method of HPV capture combined with massive paralleled sequencing (MPS). To validate the confidence of our methods, 15 cervical cancer samples were tested by PCR and the new method. Results showed that there was 100% consistence between the two methods.DNA from peripheral blood, tumor tissue, adjacent lymph nodes and adjacent normal tissue were collected from seven malignant breast cancer patients, and HPV type 16 (HPV16) was detected in 1/7, 1/7, 1/7 and 1/7 of patients respectively. Peripheral blood, tumor tissue and adjacent normal tissue were also collected from two patients with benign breast tumor, and 1/2, 2/2 and 2/2 was detected to have HPV16 DNA respectively. MPS metrics including mapping ratio, coverage, depth and SNVs were provided to characterize HPV in samples. The average coverage was 69% and 61.2% for malignant and benign samples respectively. 126 SNVs were identified in all 9 samples. The maximum number of SNVs was located in the gene of E2 and E4 among all samples. Our study not only provided an efficient method to capture HPV DNA, but detected the SNVS, coverage, SNV type and depth. The finding has provided further clue of association between HPV16 and breast cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Breast Neoplasms / virology*
  • DNA, Viral / genetics*
  • Female
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Middle Aged
  • Papillomaviridae / genetics*
  • Papillomaviridae / isolation & purification*
  • Polymorphism, Single Nucleotide / genetics
  • Uterine Cervical Neoplasms / virology

Substances

  • DNA, Viral

Grants and funding

This work was supported by the National Natural Science Fund (81272899 & 81172510) and Discipline booster plan of Xi Jing Hospital (XJZT12Z07). The study is also sponsored by Chinese 863 program (no. 2012AA02A201). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.