【摘要】 目的:探讨蛋白激酶C(protein kinase C,PKC)对大肠癌HT-29细胞间隙通讯和粘附分子表达的调节作用。方法:采用激光共聚焦对细胞间隙通讯(GJIC)功能测定,用Western blot分析E-cadherin(E-cad)、laminin receptor(LnR)、α-catenin和β-catenin粘附分子表达,研究PKC激活剂佛波酯PMA和抑制剂staurosporine(SP)对HT-29细胞间隙通讯和粘附分子表达的调节作用。结果:GJIC功能测定提示,PMA可延滞GJIC功能恢复(P<0.05)。但SP和PMA共同作用细胞后,可使GJIC功能恢复明显增快,与PMA处理组比较,差异显著(P<0.05)。Western blot分析提示,HT-29细胞可高表达E-cad、LnR,而低表达α-catenin、β-catenin。100 nmol/L PMA可诱导细胞表达LnR增强,使E-cad表达下调,但对α-catenin、β-catenin表达无影响。而SP可拮抗PMA的作用,使LnR表达下降,E-cad表达增强,对α-catenin和β-catenin的表达作用亦无影响。结论:PMA和SP对细胞表达E-cad、LnR的作用可能受到PKC调控,PMA和SP调节HT-29细胞粘附的机制可能还涉及其影响了细胞的GJIC功能,可能与PMA和SP调控细胞E-cad表达机制有关。更多还原

【Abstract】 Objective:To evaluate the regulative effects of PKC on gap junction intercellular communication(GJIC) and expression of adhesion molecules in colon carcinoma HT-29 cell.Methods:GJIC function of colon cancer HT-29 cell was detected by confocal laser scanning microscopy.Expressions of some adhesive molecules,such as E-cadherin(E-cad),laminin receptor(LnR),α-catenin and β-catenin in these cells were determined by Western blot.Regulative effects of PKC activator phorbol-12-myristate-13-acetate(PMA) and PKC inhibitor staurosporine(SP) on GJIC and expression of above adhesion molecules were investigated.Results:The determination of GJIC function of HT-29 cell indicated that its recovery rate decreased after PMA treatment(P<0.05).However,the recovery rate of GJIC function increased significantly after treatment with SP plus PMA compared with that only treated with PMA(P<0.05).The findings of Western blot showed that in HT-29 cell,the expressions of E-cad and LnR were higher than α-catenin and β-catenin.100 nmol/L PMA treatment for 4 h could induce LnR expression increasing and down regulate the E-cad expression,but it had no effects on α-catenin and β-catenin expression.SP could rivalry the effects of PMA by reducing the LnR expression and increasing E-cad expression although it also had no effects on α-catenin and β-catenin expression.Conclusion:The effects of PMA and SP on expressions of E-cad and LnR may be controlled by PKC.Mechanisms of the regulation of HT-29 cell adhesion by PMA and SP may be involved in their effects on GJIC function.These effects may be related with regulation of E-cad expression by PMA and SP.更多还原