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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 investigate the effect of PKC inhibitor phorbol ester PMA and PKC inhibitor Staurosporine (SP) on the adhesion of human colorectal cancer HT -29 cells. Methods HUVECs were detected by cell culture in vitro. The expression of E-cadherin (E Cad), Laminin receptor (LnR), α-catenin and α-catenin were detected by Western blot analysis. 9 cells adhesion effect. Results The results of in vitro cell culture and adhesion to HUVECs showed that HT-29 cells transformed from round to fibroblast-like cells after 10 0 nmol / LPMA treatment compared with the control group. The cells migrated and diffused , While the adhesion between HT-29 and HUVECs was weakened (P <0.05). However, the combination of 10 nmol / L LPMA and 100 nmol / L L of HT 2 9 cells resulted in patch formation, increased HT-29 cell adhesion, and decreased adhesion to HUVECs (P <0.05). The result of Western blot analysis indicated that the cells in control group could express E Cad and LnR at higher level and α Catenin at low level. The effect of 10 nmol / L LPA on the expression of LnR in HT-2 9 cells was enhanced, while that of E Cad was weakened slightly, which showed no effect on the expression of α-Catenin and α-Catenin. However, SP antagonized the effect of PMA, decreased the expression of LnR and enhanced the expression of E Cad, while had no effect on the expression of a-Catenin and a-Catenin. Conclusion PKC activator phorbol ester PMA can promote homogenous adhesion between tumor cells weakened, and HUVECs between?