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J.–H. Sohn, E. Bodek, P. Jha, Y. Wang, P. Bora, H. Kaplan, N. Bora; Complement Regulatory Proteins Are Important for the Survival of Human Retinoblastoma . Invest. Ophthalmol. Vis. Sci. 2006;47(13):5161.
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This study was performed to investigate the role of complement regulatory proteins – MCP, DAF and CD59 in the survival of human retinoblastoma cells.
Y79 human retinoblastoma cells (ATCC HTB 18) were obtained from the American Type Culture Collection and cultured under standard conditions using RPMI–1640 medium. The expression of CRPs was examined by flow cytometry and semi–quantative RT–PCR analysis. Monoclonal antibodies against human MCP, DAF and CD59 were used in flow cytometric analysis. Complement dependent cytotoxicity (CDC) was measured in vitro using Alamar Blue with blocking monoclonal antibody for MCP and CD59.
Our results of flow cytometry and RT–PCR analysis demonstrated that MCP and CD59 transcripts as well as proteins were abundantly expressed in retinoblastoma cells. In contrast, DAF was not expressed on these cells. Neutralization of MCP and CD59 caused increased death of retinoblastoma cells in vitro as determined by CDC assay.
Functional suppression of complement regulatory proteins in vitro results in the death of retinoblastoma cells. Thus, neutralization of complement regulatory proteins on human retinoblastoma cells may have therapeutic potential and may serve as novel therapeutic agent in future.
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