PKC激活剂佛波酯PMA和抑制剂Staurosporine对大肠癌HT-29细胞黏附作用的影响  

The Effects of Protein Kinase C Activator Phorbol-12-Myristate-13-Acetate and Inhibitor Staurosporine on Cell Adhesion of Large Bowel Carcinoma HT-29 Cells

作　　者：陈宏[1] 李荣清[1] 丁云霞[1] 林旭[1] 黄丽春[1] 刘跃[1] 张国桥[1] 张振书[2] 

机构地区：[1]成都军区昆明总医院肿瘤科,650032 [2]南方医科大学附属南方医院消化科,510515

出　　处：《实用癌症杂志》2005年第1期39-43,共5页The Practical Journal of Cancer

摘　　要：目的　探讨蛋白激酶C(PKC )激活剂佛波酯PMA和PKC抑制剂Staurosporine(SP)对大肠癌HT -2 9细胞黏附作用的影响。方法　采用体外细胞培养观察、细胞黏附人脐带静脉内皮细胞(HUVECs)测定、Westernblot分析黏附分子E Cadherin(E Cad)、LamininReceptor(LnR)、αCatenin和αCatenin表达等方法,研究PMA和SP对HT- 2 9细胞黏附作用的影响。结果　从体外细胞培养观察和黏附HUVECs实验结果发现,10 0nmol/LPMA处理后,和培养液对照组相比可见HT- 2 9细胞由圆形变成成纤维细胞样生长,细胞发生游走、扩散,HT -2 9细胞间黏附减弱,而对HUVECs的黏附增强(P <0 .0 5 )。而10 0nmol/LPMA和10 0nmol/LSP联合处理HT 2 9细胞,可见细胞成片生长,HT- 2 9细胞间黏附增强,而对HUVECs的黏附则降低(P <0 .0 5 )。Westernblot分析结果提示,培养液对照组细胞可较高水平表达E Cad、LnR ,而低水平表达αCatenin、αCatenin。10 0nmol/LPMA作用细胞可诱导HT- 2 9细胞LnR表达水平增强,而E Cad表达水平轻度减弱,对αCatenin、α- Catenin表达水平无作用。而SP可拮抗PMA的作用,使LnR表达水平降低,E Cad表达水平增强,而对α- Catenin和α- Catenin的表达亦无影响。结论　PKC激活剂佛波酯PMA可促使肿瘤细胞间的同质性黏附能力减弱,与HUVECs间?Objective To evaluate the effects of protein kinase C activator phorbol-12-myristate-13-acetate(PMA) and inhibitor staurosporine(SP) on cell adhesion of large bowel carcinoma HT-29 cells.Methods The homogeneous adhesion abilities of HT-29 cells were observed by phasecontrast microscopy and scanning electron microscopy,the heterogeneous adhesion were studied by means of an adherent to human umbilical vein endothelial cells(HUVECs) model.Some types of adhesive molecules(E-cadherin(E-Cad),laminin receptor(LnR),α-catenin and α-catenin) expressions of cancer cells were analysised by western blot assay.Results 100 nmol/L PMA treatment for 4 hours could induce cellular morphological changes from round shape to fibroblast-like shape,the ability to shift outside from the cell-island was found.The ability of cell-cell adhesion was reduced and cell-HUVECs adhesion was increased(P<0.05).However,100 nmol/L SP plus 100 nmol/L PMA treatment could induce the ability of cell-cell adhesion to increase and cell-HUVECs adhesion to decrease(P<0.05).The expressions of E-Cad and LnR were much high than α-catenin and α-catenin in HT-29 cell.100 nmol/L PMA treatment for 4 hours could induce LnR expression to increase and induce the E-Cad expression to down-adjust lightly,but no effects on α-catenin and α-catenin expressions.However,SP could inhibit the effects of PMA and induce the LnR expression to decrease markedly and the E-Cad expression to up-adjust obviously,but also no clear effects on adjusting the expressions of α-catenin and α-catenin.Conclusion PMA could decrease the ability of homogeneous adhesion of HT-29 cells and increase the ability of heterogeneous adhesion of cell-HUVECs,PMA could also increase the LnR expression and decrease the E-Cad expresson of HT-29 cells.However,SP could inhibit the effects induced by PMA and induces reverse effects.The results speculate that the homogeneous adhesion and heterogeneous adhesion of HT-29 cells may be mediated by adhesive molecules E-Cad and LnR,respectively.The above eff

关 键 词：STAUROSPORINE 细胞黏附作用 PMA 佛波酯 激活剂 大肠癌 Catenin HT-29细胞 蛋白激酶C(PKC) HUVECs Western 人脐带静脉内皮细胞 体外细胞培养 Cad表达 Laminin PKC抑制剂 blot 黏附能力 LnR 低水平表达 肿瘤细胞间 黏附分子 

分 类 号：R979.1[医药卫生—药品] R87[医药卫生—药学]