Detection of breast cancer micrometastases in axillary lymph nodes by means of reverse transcriptase-polymerase chain reaction. Comparison between MUC1 mRNA and keratin 19 mRNA amplification

Am J Pathol. 1996 Feb;148(2):649-56.

Abstract

Usefulness of MUC1 mRNA and keratin 19 mRNA as a target of reverse-transcriptase polymerase chain reaction (RT-PCR) was compared in the detection of breast cancer micrometastases in axillary lymph nodes. RT-PCR amplification of MUC1 mRNA and keratin 19 mRNA was conducted using total RNA samples. RT-PCR products were stained with ethidium bromide and analyzed by agarose gel electrophoresis. Expression of both MUC1 mRNA and keratin 19 mRNA was detected by RT-PCR in a breast cancer cell line (MRK) and in all the 23 primary breast cancers but not in the control lymph nodes obtained from patients with benign diseases. A serial dilution study of MRK cells against normal lymph node cells has shown that detection sensitivity of MUC1 RT-PCR and keratin 19 RT-PCR were 1/10(5) and 1/10(6) (cancer/lymph node cells), respectively. Sixty-three axillary lymph nodes were obtained from 23 patients with primary breast cancer, and metastases in each lymph node were investigated by histological examination (hematoxylin and eosin sections) and RT-PCR method. In all 10 lymph nodes, which were histologically metastasis-positive, both MUC1 mRNA and keratin mRNA were detected by RT-PCR. Of the 53 histologically negative lymph nodes, 3 (6%) and 5 (9%) lymph nodes were found to express MUC1 mRNA and keratin 19 mRNA, respectively, indicating the presence of micrometastases which could be detected by RT-PCR but not by histological examination. These results demonstrate the usefulness of both MUC1 RT-PCR and keratin 19 RT-PCR in the detection of breast cancer micrometastases in lymph nodes, and also indicate the superiority of keratin 19 RT-PCR over MUC1 RT-PCR because of its higher detection sensitivity.

Publication types

  • Comparative Study

MeSH terms

  • Axilla
  • Base Sequence
  • Breast Neoplasms / pathology*
  • DNA Primers
  • Female
  • Humans
  • Keratins / analysis*
  • Keratins / genetics
  • Lymph Nodes / chemistry
  • Lymph Nodes / pathology
  • Lymphatic Metastasis / diagnosis*
  • Lymphatic Metastasis / pathology
  • Molecular Sequence Data
  • Mucin-1 / analysis*
  • Mucin-1 / genetics
  • Polymerase Chain Reaction / methods*
  • RNA, Messenger / analysis*
  • RNA, Messenger / genetics
  • RNA-Directed DNA Polymerase
  • Retrospective Studies
  • Sensitivity and Specificity

Substances

  • DNA Primers
  • Mucin-1
  • RNA, Messenger
  • Keratins
  • RNA-Directed DNA Polymerase