Localization of thymidine phosphorylase expression in colorectal carcinoma tissues by in situ RT-PCR assay.
Abundant expression of thymidine phosphorylase (TP) mRNA and protein was observed in human carcinoma tissues relative to the corresponding normal tissues. However, the distribution of cells that express TP has not yet been determined. In this study, we examined enzyme activity of TP in 24 primary colorectal carcinoma tissues and the corresponding normal tissue. Using in situ reverse-transcription polymerase chain reaction (RT-PCR), a new methodology to determine the localization of genes of interest objectively and clearly, cell distribution was compared. The expression of TP mRNA is observed in normal infiltrating cells adjacent to colon carcinoma cells by in situ RT-PCR. Similar to the protein activity, the average TP mRNA expression (amplified products/x400 field) in cases of Dukes C and D is 76.8, while that of Dukes A and B is 42.5, being significantly different (p = 0.001). Moreover, the number of amplified products in colon carcinoma tissues showed a significant correlation with the enzyme activity evaluated by high-performance liquid chromatography (p < 0.01, R = 0.76). In conclusion, the expression of TP mRNA is localized in stromal tissues as determined by in situ RT-PCR and can reflect the level of TP enzyme activity.
Department of Surgery, Medical Institute of Bioregulation, Kyushu University, Beppu, Japan.
Koshi
K
Mimori
Ayumi
A
Matsuyama
Keiji
K
Yoshinaga
Keishi
K
Yamashita
Takaaki
T
Masuda
Hiroshi
H
Inoue
Hiroaki
H
Ueo
Masaki
M
Mori
D000230:Adenocarcinoma / Q000201:enzymology*; D002288:Adenocarcinoma, Mucinous / Q000201:enzymology* / Q000473:pathology; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002454:Cell Differentiation; D002851:Chromatography, High Pressure Liquid; D015179:Colorectal Neoplasms / Q000201:enzymology* / Q000473:pathology; D017931:DNA Primers / Q000737:chemistry; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009367:Neoplasm Staging; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013939:Thymidine Phosphorylase / Q000235:genetics* / Q000378:metabolism; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D012333:RNA, Messenger; D013939:Thymidine Phosphorylase