Huang Ruochun, Lin Ying, Wang Cheng C, Gano Jacob, Lin Biaoyang, Shi Qian, Boynton Alton, Burke Jocelyn, Huang Ruo-Pan
Department of Gynecology and Obstetrics, Emory University School of Medicine, 1639 Pierce Drive, Atlanta, GA 30322, USA.
Cancer Res. 2002 May 15;62(10):2806-12.
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.
此前，我们证明了连接蛋白43（cx43）可抑制人胶质母细胞瘤细胞的生长。为了研究cx43抑制肿瘤的分子机制，我们开发了一种人细胞因子阵列系统，该系统可同时检测43种细胞因子的表达。通过使用这项新技术，我们分析了cx43转染细胞中cx43调控的基因。细胞因子阵列显示，cx43转染细胞中单核细胞趋化蛋白-1（MCP-1）的表达显著降低。逆转录-聚合酶链反应（RT-PCR）、免疫蛋白质印迹和cDNA微阵列进一步证实了这一观察结果。将对照转染细胞的条件培养基和重组MCP-1添加到cx43转染细胞中，可显著增强cx43转染细胞在软琼脂中的增殖和集落形成。相反，添加抗MCP-1中和抗体可显著抑制对照转染细胞的增殖。我们的结果表明，MCP-1参与了cx43对人胶质母细胞瘤细胞生长的抑制作用。
