The hyaluronan receptor RHAMM/IHABP in astrocytoma cells: expression of a tumor-specific variant and association with microtubules.

Hyaluronan binding to its cellular receptors CD44 and ICAM-1 appears to enhance the malignant behavior of tumors, including astrocytomas. RHAMM/IHABP, another hyaluronan receptor, has been identified in breast carcinoma cells, but its presence in astrocytomas is yet undetermined. Herein, we report that a monoclonal antibody against plectin (a cytoskeletal protein linker) recognizes on Western blots of U-373 MG glioblastoma cells, a 300-kDa band corresponding to plectin and two bands of 86 and 70 kDa. cDNA cloning and Northern blotting reveals that these two bands represent isoforms of RHAMM/IHABP. Sequence comparisons suggest that the plectin monoclonal antibody recognizes RHAMM/IHABP because this protein and plectin share short peptide sequences of similar primary and secondary structure. Western blotting demonstrates that most human astrocytoma tissues and cell lines express the 86- and 70-kDa isoforms of RHAMM/IHABP. Interestingly, the 70-kDa variant is undetectable in normal brain tissues and in primary cultures of astrocytes suggesting that its expression is tumor-specific. Transfection experiments with epitope-tagged RHAMM/IHABP cDNA established that RHAMM/IHABP associates with microtubules in astrocytoma cells, while in normal astrocytes it either co-localizes with microtubules or has a diffuse cytoplasmic distribution. This suggests that RHAMM/IHABP has the capacity to bind to microtubules in normal and transformed astrocytes, and that neoplasia may favor this association.