Role of gap junctional intercellular communication on the cytotoxic effect of anticancer drug

我们研究了间隙连接细胞间通讯（GJIC）在抗癌药物多西紫杉醇对食管癌细胞系细胞毒性作用中的作用。RT-PCR和Western blot分析显示，人食管癌细胞株KE-10不表达任何连接蛋白（connexin, Cx）和GJIC能力。将Cx32基因转染到细胞中。转染Cx32的细胞（KE-10/Cx32）经RT-PCR、Western blot和免疫组织化学检测证实Cx32表达。染料转移实验显示，KE-10/Cx32恢复了GJIC能力，细胞毒性比KE-10增强了30-40%。KE-10/C32细胞凋亡上调。而导致多西紫杉醇耐药的MDR基因在两种细胞系中均未表达。最后，我们研究了GJIC抑制剂卡贝诺洛酮对多西紫杉醇细胞毒作用的影响。卡贝诺洛酮使多西紫杉醇对KE-10/Cx32细胞的细胞毒性降低20%。这些数据表明GJIC参与了多西紫杉醇的细胞毒性作用。

We investigate the role of gap junctional intercellular communication(GJIC)on the cytotoxic effect of an anticancer drug, docetaxel, in esophageal cancer cell line.RT-PCR and Western blot analysis revealed that human esophageal cancer cell line, KE-10, didn't have expression of any connexin (Cx) and GJIC capacity. Cx32 gene was transfected into the cells. Cx32 expression was confirmed with RT-PCR, Western blot and immunohistochemisty in the Cx32-transfected cells (KE-10/Cx32). KE-10/Cx32 showed the restoration of GJIC capacity with dye transfer assay and the cyotoxicity was enhanced by 30-40% compared with KE-10. Apoptosis was upregulated in KE-10/C32 cells. And MDR gene, which is responsible for the drug resistance of docetaxel, wasn't expressed in two cell lines. Finally, we investigated the effect of the GJIC inhibitor, carbenoxolone, on cytotoxic effect of docetaxel. Carbenoxolone decreased the cytotoxity of docetaxel in KE-10/Cx32 cells by 20%. These date suggest that GJIC is involved in the cytotoxic effect of docetaxel.