Two initiation sites of early detection of colon cancer revealed by localization of pERK1/2 in the nuclei or in aggregates at the perinuclear region of the tumor cells.
We have used human specimens and antibodies to pERK1/2 to detect early development of colon cancer using indirect immunocytochemistry. Two distinct sites were stained; one at the tip of the colon crypts and the other in the stromal tissue associated with the colonic tissue. These foci represent early stages of colon cancer initiation sites as established by enhanced Kirsten Rat Sarcoma Virus (KRAS) and the lack of p53 staining. The enhanced KRAS coincides with the initiation of tumor growth revealed by pERK1/2, both in the tip of the colon crypts, as well as in the stromal initiation site of the colon tumors. Foci of pERK1/2 staining were also detected in 50% of stromal tissue and tips of colon crypts, which were classified as normal tissues, adjacent to the malignant tissue according to general morphology. However, in colon specimens, where no malignancy was observed, no accumulation of pERK1/2 was observed. The staining of pERK1/2 at the stromal foci of the apparently non-malignant tissue appeared as aggregates in the perinuclear region, while in the colon epithelium it appeared in the cell nuclei. In low-grade colon cancer that was still free of induced mutated p53, staining of pERK1/2 was prominent in the cell nuclei, both in the stroma tissue and the tip of the colon crypts. In the intermediate stage, that exhibited significant p53 staining, only a fraction of p53-free tumor cells was labeled with pERK1/2 antibody, while in high-grade tumors, all cells of tumors were labeled with antibodies to p53, but not with antibodies to pERK1/2. We suggest that the down regulation in pERK1/2 labeling is due to the mitogenic capacity of the tumor cells, which are shifted from being driven by nuclear pERK1/2 to mutated p53 expression. We also found that the cytoplasm of low grade tumors was positive for epiregulin, while this labeling decreased in high-grade tumors. We found that the tumors arising from the stroma demonstrated poor structural differentiation, while the tumors initiating from the epithelial cells of the colon demonstrated high structural differentiation. We conclude that pERK1/2 is a sensitive marker of early colon cancer, which disappears at later stages of cancer development. Moreover, pERK1/2 staining can distinguish between tumor cells originating from the tip of the colon crypts and those developing in the stroma, which is present in the close vicinity to colon epithelial tissue, and thus can assist in selecting the appropriate therapy.
Department of Molecular Cell Biology, The Weizmann Institute of Science, Rehovot 76100, Israel. abraham.amsterdam@weizmann.ac.il
Abraham
A
Amsterdam
Elias
E
Shezen
Calanit
C
Raanan
Letizia
L
Schreiber
Yasmin
Y
Slilat
Yakov
Y
Fabrikant
Ehud
E
Melzer
Rony
R
Seger
D000230:Adenocarcinoma / Q000175:diagnosis* / Q000201:enzymology; D014408:Biomarkers, Tumor / Q000378:metabolism*; D002467:Cell Nucleus / Q000201:enzymology*; D003110:Colonic Neoplasms / Q000175:diagnosis* / Q000201:enzymology; D042241:Early Diagnosis; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D020935:MAP Kinase Signaling System / Q000502:physiology; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D009367:Neoplasm Staging; D017154:Stromal Cells / Q000201:enzymology / Q000473:pathology
D014408:Biomarkers, Tumor; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3
Cancer stem cells; Colon cancer; Epiregulin; KRAS; MAPK; p53; pERK1/2