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.
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.
Noguchi
S
S
Department of Surgery, Center for Adult Diseases, Osaka, Japan.
Aihara
T
T
Motomura
K
K
Inaji
H
H
Imaoka
S
S
Koyama
H
H
D001365:Axilla; D001483:Base Sequence; D001943:Breast Neoplasms / Q000473:pathology*; D017931:DNA Primers; D005260:Female; D006801:Humans; D007633:Keratins / Q000032:analysis* / Q000235:genetics; D008198:Lymph Nodes / Q000737:chemistry / Q000473:pathology; D008207:Lymphatic Metastasis / Q000175:diagnosis* / Q000473:pathology; D008969:Molecular Sequence Data; D018396:Mucin-1 / Q000032:analysis* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D012194:RNA-Directed DNA Polymerase; D012189:Retrospective Studies; D012680:Sensitivity and Specificity
D017931:DNA Primers; D018396:Mucin-1; D012333:RNA, Messenger; D007633:Keratins; D012194:RNA-Directed DNA Polymerase