Characterization of human PA2.26 antigen (T1α-2, podoplanin), a small membrane mucin induced in oral squamous cell carcinomas

Abstract

We report the full cDNA sequence encoding the human homologue of murine PA2.26 (T1␣-2, podoplanin), a small mucin-type transmembrane glycoprotein originally identified as a cell-surface antigen induced in keratinocytes during mouse skin carcinogenesis. The human PA2.26 gene is expressed as 2 transcripts of 0.9 and 2.7 kb in several normal tissues, such as the placenta, skeletal muscle, heart and lung. Using a specific polyclonal antibody raised against a synthetic peptide of the protein ectodomain, PA2.26 was immunohistochemically detected in about 25% (15/61) of human early oral squamous cell carcinomas. PA2.26 distribution in the tumours was heterogeneous and often restricted to the invasive front. Double immunofluorescence and confocal microscopy analysis showed that PA2.26 colocalized with the membrane cytoskeleton linker ezrin at the surface of tumour cells and that its presence in vivo was associated with downregulation of membrane E-cadherin protein expression. Ectopic expression of human PA2.26 in HeLa carcinoma cells and immortalized HaCaT keratinocytes promoted a redistribution of ezrin to the cell edges, the formation of cell-surface protrusions and reduced Ca 2؉ -dependent cell-cell adhesiveness. These results point to PA2.26 as a novel biomarker for oral squamous cell carcinomas that might be involved in migration/invasion.Key words: mucin; PA2.26; ezrin; E-cadherin; microvilli; OSCC Squamous cell carcinomas (SCCs) of the oral cavity, pharynx and larynx remain a significant public health problem. They represent 2-3% of all malignancies, and their incidence, particularly that of oral SCCs (OSCCs), is increasing in Western countries. 1 In spite of improved therapeutic procedures, the prognosis of OSCC patients remains poor and considerably lower than that of other neoplasias. 2 This fact can be attributed to several factors: failure to respond to available chemotherapy, late presentation of the lesions and lack of suitable markers for early detection and prognosis. 3,4 Hence, the finding of novel tumour markers, particularly those associated with tumour cell invasion and spreading, can help provide a more accurate evaluation of prognosis and a more efficient management of the disease.PA2.26 antigen was identified in our laboratory as a cell-surface protein induced in murine epidermal keratinocytes and dermal fibroblast-like cells during wound healing and chemical carcinogenesis. 5 Sequence analysis of the isolated cDNA and biochemical characterization of the protein revealed that murine PA2.26 is a small mucin-like transmembrane glycoprotein of about 45 kDa, 6 highly homologous to the rat alveolar type I cell marker T1␣ and the podocyte-associated glycoprotein podoplanin. 7,8 Murine PA2.26 nucleotide sequence is almost identical to that of OTS-8 and gp38, markers of the osteoblastic cell lineage and stromal cells in peripheral lymphoid tissues, respectively, 9,10 and completely matches the nucleotide sequence of RANDAM-2, a recently discovered membrane glycoprotein expressed in neuronal cells during m...

Podoplanin expression correlates with sentinel lymph node metastasis in early squamous cell carcinomas of the oral cavity and oropharynx

“…[15][16][17][18][19][20][21] These findings suggest that podoplanin plays a role in the progression of epithelial cancer. However, the exact molecular function of cancer cell-expressed podoplanin is currently studied.…”

Low podoplanin expression in pretreatment biopsy material predicts poor prognosis in advanced-stage squamous cell carcinoma of the uterine cervix treated by primary radiation

“…23 In vitro studies suggested that podoplanin may be involved in mediating cell motility and adhesion. 23,24 Very recently, podoplanin expression was reported in squamous cell carcinomas of the epidermis, 13 oral cavity, 24 and lung, 25 and it was suggested that it may be involved in tumor cell invasion, metastasis, and tumor progression. In addition, mucin-type glycoproteins, such as podoplanin, are expressed widely on human normal cells and tumors [26][27][28] and have recently been implicated in cellular signaling, cell adhesion, tumor growth, and suppression of apoptosis.…”

The potential role of podoplanin in tumour invasion

“…The extracellular domain is highly O-glycosylated, with sialic acid, a-2,3 linked to galactose, forming the main part of the protein's carbohydrate moieties. In normal human tissue, podoplanin is expressed in kidney podocytes (Breiteneder-Geleff et al, 1999), in skeletal muscle, placenta, lung and heart (Martin-Villar et al, 2005), in myofibroblasts of the breast and salivary glands, in osteoblasts and mesothelial cells (Ordonez, 2006b). It is also expressed on the apical surface of rat alveolar type I cells (Rishi et al, 1995).…”

“…The expression of podoplanin is upregulated in a number of different human cancers, including squamous cell carcinoma of the oral cavity, the larynx, the lung, the cervix, the oesophagus, and the skin, in dysgerminomas of the ovary and granulosa cell tumours, in mesothelioma, and in many tumours of the central nervous system (CNS) (Kato et al, 2005;Kimura and Kimura, 2005;Martin-Villar et al, 2005;Schacht et al, 2005;Shibahara et al, 2006;Wicki et al, 2006). The oncofetal M2A antigen expressed in testicular germ cell tumours is identical to podoplanin (Dumoff et al, 2005).…”