PTEN͞MMAC1 is a tumor suppressor gene located on chromosome 10q23. Inherited PTEN͞MMAC1 mutations are associated with a cancer predisposition syndrome known as Cowden's disease. Somatic mutation of PTEN has been found in a number of malignancies, including glioblastoma, melanoma, and carcinoma of the prostate and endometrium. The protein product (PTEN) encodes a dualspecificity protein phosphatase and in addition can dephosphorylate certain lipid substrates. Herein, we show that PTEN protein induces a G 1 block when reconstituted in PTEN-null cells. A PTEN mutant associated with Cowden's disease (PTEN;G129E) has protein phosphatase activity yet is defective in dephosphorylating inositol 1,3,4,5-tetrakisphosphate in vitro and fails to arrest cells in G 1 . These data suggest a link between induction of a cell-cycle block by PTEN and its ability to dephosphorylate, in vivo, phosphatidylinositol 3,4,5-trisphosphate. In keeping with this notion, PTEN can inhibit the phosphatidylinositol 3,4,5-trisphosphate-dependent Akt kinase, a downstream target of phosphatidylinositol 3-kinase, and constitutively active, but not wild-type, Akt overrides a PTEN G 1 arrest. Finally, tumor cells lacking PTEN contain high levels of activated Akt, suggesting that PTEN is necessary for the appropriate regulation of the phosphatidylinositol 3-kinase͞Akt pathway.

We have previously demonstrated a link between alpha2,6-Sialyltransferase (alpha2,6-ST; E.C. 2.4.99.1) expression and differentiation of colon tumors. So far, information is not available relative to the expression of alpha2,6-ST in tumors and the survival of patients with colorectal cancer. We have examined the expression of alpha2,6-ST in a variety of colorectal adenocarcinomas (n = 46) at different stages of differentiation (G1 to G3) by immunoperoxidase assay using monoclonal antibody (MAb) 6B9. Clinical outcome of the patients in a 5-year follow-up study has been correlated with the expression of alpha2,6-ST in tumors surgically removed from the same patients. No significant difference in the alpha2,6-ST expression was noted when age, sex, and tumor locations (colon, rectum) were included as parameters. However, 52% of the moderate (G2) and well-differentiated (G1) adenocarcinomas showed stronger alpha2,6-ST expression compared with poorly differentiated (G3) adenocarcinomas. Notably, absence to moderate levels of tumor alpha2,6-ST expression was correlated with 100% survival in patients with stage I and II tumors compared with 64% survival in patients with strong tumor alpha2,6-ST expression (p < 0.01). These studies suggest a negative correlation between the expression of alpha2,6-ST in tumors and a good clinical outcome in colorectal cancer patients.

It has been reported that alpha2,6-sialyltransferase (alpha2,6-ST; E.C. 2.4.99.1) activity is associated with cellular differentiation. To define its role in colon carcinoma differentiation, we have generated murine monoclonal antibodies (MAb) against alpha2,6-sialyltransferase. The MAb, designated 6B9 of IgM isotype, showed strong reactivity with the purified and crude alpha2,6-ST by ELISA and dot blot assays. Western blotting with MAb 6B9 identified purified alpha2,6-ST of MW 47 kDa and the same MW protein from rat and human liver extracts. The MAb also reacted with two other liver proteins of approximate MW 65 and 100 kDa. Immunoperoxidase studies with formalin-fixed paraffin-embedded tissues showed that MAb 6B9 reacts with liver tissues, the staining of hepatocytes was granular and cytoplasmic. There was a distinct pattern of zonal distribution of this enzyme in hepatocytes located particularly in the portal areas of the liver corresponding to zone 1. Normal colon (100%) and hyperplastic polyps (100%) showed very weak to no reactivity. Adenomas (100%) demonstrated moderate reactivity, while the poor (33%), moderate (100%) and well-differentiated (80%) colon adenocarcinomas showed strong reactivity. Results suggest that alpha2,6-ST is associated with the differentiation state of colon tumors.