Quantification of micrometastases in lymph nodes of colorectal cancer using real-time fluorescence polymerase chain reaction.
The expression of carcinoembryonic antigen (CEA) mRNA was assessed in 102 lymph nodes (LNs) obtained from seven colorectal cancer patients by both the conventional non-quantitative RT-PCR and quantitative RT-PCR. The number of CEA-expressing cells was calculated compared with CEA-expressing MKN-45 cell line as a standard control. Using the quantitative RT-PCR, the relative number of CEA-expressing cells ranged between 1.3x103 and 5.7x106 in 16 histologically positive LNs and between 2.3x101 and 8.1x105 in 10 histologically negative and RT-PCR positive LNs. In both histologically and RT-PCR negative LNs, the relative cell number was <4.0x102. Our results demonstrated that quantifying the amount of metastasis might enhance the reliability of RT-PCR detection assay as a diagnostic tool for the detection of cancer micrometastases.
Miyake
Y
Y
Department of Surgery II, Osaka University Medical School, Osaka 565-0871, Japan.
Fujiwara
Y
Y
Ohue
M
M
Yamamoto
H
H
Sugita
Y
Y
Tomita
N
N
Sekimoto
M
M
Shiozaki
H
H
Monden
M
M
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D052160:Benzothiazoles; D002272:Carcinoembryonic Antigen / Q000032:analysis*; D015179:Colorectal Neoplasms / Q000737:chemistry* / Q000473:pathology*; D003959:Diamines; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D008198:Lymph Nodes / Q000737:chemistry*; D008207:Lymphatic Metastasis; D008297:Male; D008875:Middle Aged; D009930:Organic Chemicals; D011804:Quinolines; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D052160:Benzothiazoles; D002272:Carcinoembryonic Antigen; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I