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Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 2001 Jul;54(7):533–538. doi: 10.1136/jcp.54.7.533

Expression of transcription factor AP-2 in colorectal adenomas and adenocarcinomas; comparison of immunohistochemistry and in situ hybridisation

K Ropponen 1, J Kellokoski 1, R Pirinen 1, K Moisio 1, M Eskelinen 1, E Alhava 1, V Kosma 1
PMCID: PMC1731475  PMID: 11429425

Abstract

Aims—To investigate whether the three different AP-2 isoforms are expressed differently in colorectal adenomas and carcinomas.

Methods—The study comprised 43 randomly selected patients diagnosed and treated at Kuopio University Hospital in 1996 for colorectal adenocarcinoma (n = 30) and colorectal adenoma (n = 13). The expression of AP-2α, AP-2ß, and AP-2γ was analysed by immunohistochemistry (IHC) and the mRNA status of AP-2α was determined by in situ hybridisation (ISH) and confirmed by reverse transcription polymerase chain reaction (RT-PCR). AP-2 expression patterns were correlated with clinicopathological variables.

Results—In adenomas and carcinomas, AP-2ß cytoplasmic positivity was higher than that of AP-2α or AP-2γ. AP-2α expression was reduced in advanced Dukes's stage carcinomas. In high grade carcinomas, both AP-2α and AP-2γ expression was reduced. ISH demonstrated increased AP-2α values in high grade carcinomas. Seven of 30 carcinoma specimens displayed a moderate or strong mRNA signal, despite being negative for AP-2α protein. RT-PCR from AP-2α mRNA and protein positive tumours confirmed that the positive signal in ISH originated from the exon 2 of TFAP2A.

Conclusions—AP-2α was reduced in advanced Dukes's stage adenocarcinomas. Together with reduced AP-2γ expression in high grade carcinomas, this might contribute to tumour progression. The discrepancy between mRNA and protein expression suggests that post-transcriptional regulatory mechanisms might modify the availability of functional AP-2α protein in colorectal carcinoma.

Key Words: AP-2 proteins • immunohistochemistry • in situ hybridisation • colorectal neoplasms

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Figure 1 AP-2 immunohistochemical analysis: diffuse cytoplasmic (A) AP-2α, (B) AP-2ß, and (C) AP-2γ expression in colorectal adenocarcinoma cells. Bar, 60 µm.

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Figure 2 (A) mRNA in situ hybridisation analysis showing strong diffuse (+++) AP-2α expression in colorectal adenocarcinoma and (B) negative control. Bar, 60 µm.

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Figure 3 Reverse transcription polymerase chain reaction (RT-PCR) analysis of AP-2α expression was performed on the following extracts: lanes 1 and 6, x174 DNA/Hin FI marker (0.4 µg); lane 2, a negative PCR control for ß actin in which the template was replaced by water; lane 3, a positive control for ß actin RNA (from Clontech human placenta cDNA library); lane 4, case number 28 contains a PCR product of the same molecular size (156 bp; ß actin) as the positive control sample; lane 5, case number 28 contains a PCR product of the same molecular size (132 bp; AP-2α) as the positive control sample; lane 7, positive control sample (from HeLa cDNA; AP-2α); lane 8, a negative PCR control for AP-2α in which the template was replaced by water.

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