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Connexin 43 Suppresses Human Glioblastoma Cell Growth by Down-Regulation of Monocyte Chemotactic Protein 1, as Discovered Using Protein Array Technology¹

Department of Gynecology and Obstetrics, Emory University School of Medicine, Atlanta, Georgia 30322 [R. H., Y. L., Q. S., R-P. H.]; Packard Bioscience, Meriden, Connecticut 06450 [C. C. W., J. B.]; Northwest Biotherapeutics, Inc., Bothell, Washington [J. G., A. B.]; and Institute for Systems Biology, Seattle, Washington 98105-6099 [B. L.]

Previously, we demonstrated that connexin 43 (cx43) suppressed the growth of human glioblastoma cells. To investigate the molecular mechanisms involved in tumor suppression by cx43, we developed a human cytokine array system, which simultaneously detects the expression of 43 cytokines. By using this new technology, we analyzed the cx43-regulated genes in cx43-transfected cells. The cytokine arrays showed that expression of monocyte chemotactic protein-1 (MCP-1) was profoundly reduced in cx43-transfected cells. RT-PCR, immuno-Western blot, and cDNA microarrays further confirmed this observation. Addition of conditioned medium from control-transfected cells and recombinant MCP-1 to cx43-transfected cells significantly enhanced cx43-transfected cell proliferation and colony formation in soft agar. In contrast, addition of neutralization antibody against MCP-1 significantly inhibited cell proliferation in control-transfected cells. Our results suggested that MCP-1 is involved in the suppression of human glioblastoma cell growth by cx43.

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