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CD24 is a potential oncogene reported to be overexpressed in a large variety of human malignancies. We have shown that CD24 is overexpressed in 90% of colorectal tumors at a fairly early stage in the multistep process of carcinogenesis. Anti-CD24 monoclonal antibodies (mAb) induce a significant growth inhibition in colorectal and pancreatic cancer cell lines that express the protein. This study is designed to investigate further the effects of CD24 down-regulation using mAb or small interfering RNA in vitro and in vivo. Western blot analysis showed that anti-CD24 mAb induced CD24 protein down-regulation through lysosomal degradation. mAb augmented growth inhibition in combination with five classic chemotherapies. Xenograft models in vivo showed that tumor growth was significantly reduced in mAb-treated mice. Similarly, stable growth inhibition of cancer cell lines was achieved by down-regulation of CD24 expression using short hairpin RNA (shRNA). The produced clones proliferated more slowly, reached lower saturation densities, and showed impaired motility. Most importantly, down-regulation of CD24 retarded tumorigenicity of human cancer cell lines in nude mice. Microarray analysis revealed a similar pattern of gene expression alterations when cells were subjected to anti-CD24 mAb or shRNA. Genes in the Ras pathway, mitogen-activated protein kinase, or BCL-2 family and others of oncogenic association were frequently down-regulated. As a putative new oncogene that is overexpressed in gastrointestinal malignancies early in the carcinogenesis process, CD24 is a potential target for early intervention in the prevention and treatment of cancer.

CD24 was first described in the early 1980s and only attributed to scattered publications, referred to as a cell surface molecule in hematopoiesis. Recently, studies are accumulating to show that CD24 conveys a function in cell-to-cell interaction and regulation of proliferation and adhesion. CD24 appears to be highly expressed in a large variety of human cancers and to contribute to the acceleration of tumor growth and metastases shedding by binding to platelet (P)-selectin, L1 and by evoking--to date unknown--intracellular signal pathways. Anti-CD24 monoclonal antibodies thus act as a promising cancer treatment as was shown in the setting of gastrointestinal cancers. Recent articles also correlate CD24 expression with the identification of 'tumor stem cells'.

Background & Aims: The aim of this study was to identify genes that play a role in colorectal cancer (CRC) carcinogenesis by analysis of differential gene expression of normal and transformed CRC cell lines., Methods: Gene expression array analysis ([RG-U34] GeneChip) was performed in normal and transformed rat intestinal epithelial cells before and after exposures to celecoxib. In particular, we were looking for (1) altered gene expression in the transformed cells that reverts to normal following exposure to a selective cyclooxygenase-2 inhibitor, (2) novel genes, and (3) genes encoding membrane receptors or ligands. As a validation of the results and for human patients, immunohistochemistry was performed on 398 biological samples from the gastrointestinal tract (normal, polyps, and adenocarcinomas). Human cancer cell lines were tested for their response to anti-CD24 monoclonal antibodies., Results: A total of 1081 genes were differently expressed following malignant transformation; 71 genes showed altered expression that reverted to normal following treatment with celecoxib, including the CD24 gene. Immunohistochemistry confirmed that increased expression of CD24 is an early event in CRC carcinogenesis. It was expressed in 90.7% of adenomas and 86.3% of CRCs. Very low expression was seen in normal epithelium (16.6%). Human cancer cell lines showed growth inhibition in response to the antibodies, according to their expression levels of CD24 and in dose- and time-dependent manners. These results were repetitive for 3 different antibodies., Conclusions: CD24 is overexpressed in the colonic mucosa, already at an early stage of carcinogenesis. It may be a useful target for early detection and in CRC therapy.

Patients with pancreatic adenocarcinoma have a doom prognosis these tumors were previously proved to express high level of CD24. The current study was aimed to demonstrate that the treatment with monoclonal antibodies to CD24 is effective, in vitro, in pancreatic cancer cells, similar to what we had previously shown in the setting of colorectal cancer. Three human pancreatic cancer cell lines, Colo357, Panc1 and MIA-PaCa, were analyzed for their expression levels of CD24 by Western blot analysis. The correlation for the protein available on the cytoplasmic membrane was assessed by ELISA assay to plates coated with fixed cells using anti-CD24 Ab as the first binder. Human cancer cell lines were tested for their response to two different anti-CD24 monoclonal antibodies and a control antibody (mouse anti-GFP). Human pancreatic adenocarcinomas cell lines that express CD24 (Colo357 and Panc1 cells) showed growth inhibition in dose and time dependent manners. These results were repetitive for the two different antibodies. Growth rate was not affected in MIA-PaCa cells that do not express CD24, or when cells were treated with a control antibody. CD24 may play an important role in the carcinogenesis process of pancreatic cancer. It may serve as a useful target in the therapy of pancreatic cancer.