摘要
目的:探讨蛋白酶活化受体1(PAR1)在凝血酶促进肺癌细胞迁移、黏附、克隆形成及胶原收缩中的作用。方法:将本实验室已构建的重组干扰载体pSilence-shPAR1和过表达载体pIRES-EGFP-PAR1分别转入人肺癌细胞PLA-801D和PLA-801C中,采用反转录PCR和实时荧光定量PCR方法检测细胞中PAR1的表达量,Transwell实验检测细胞的迁移能力,细胞黏附实验检测细胞对细胞外基质的黏附能力,克隆形成实验检测细胞的增殖能力,胶原收缩实验评价细胞对细胞外基质的重塑能力。结果:PAR1高表达(PLA-801C-pIRES-EGFP-PAR1)可明显增强PLA-801C细胞的迁移、黏附和胶原收缩能力(P<0.05,P<0.001),而PAR1被有效干扰后(PLA-801D-pSi lence-shPAR1),PLA-801D细胞的迁移、黏附和胶原收缩能力显著减弱(P<0.05,P<0.001),而且PAR1的表达量直接与PLA-801D和PLA-801C细胞的克隆形成能力呈正相关。结论:PAR1在凝血酶促进肺癌细胞的黏附、迁移增殖和细胞外基质重塑等功能中发挥着重要作用,为以PAR1为靶的抗肿瘤治疗提供了理论基础。
Objective： To evaluate the role of protease-activated receptor 1 （PAR1） in thrombin-mediated lung cancer cell function responses. Methods： The recombinant vector pSilence-shPAR1 and pIRES-EGFP-PAR1 plas- raids were transferred into lung cancer cell PLA-gO1D and PLA-801C, respectively. Reverse transcription PCR and real-time fluorescent quantitative PCR were used to detect the level of PAR1 mRNA. Transwell migration chamber was used to evaluate the ability of migration. The ability of the cell adhesion to extracellular matrix was evaluated with cell adhesion assay. The proliferation of the cell was measured with colony formation assay. The ability of collagen remodeling by lung cancer cells was measured with collagen contraction assay. Results： The overexpression of PAR1 in PLA-801C cells（PLA-801C-pIRES-EGFP-PAR1） significantly enhanced thrombin-mediated cell migration, adhesion, proliferation and collagen contraction（P〈0.05, P〈0.001）. The interference of PAR1 in PLA-801D cells （PLA-801D-pSilenee-shPAR） markedly attenuated the thrombin-mediated cell function （P〈0.05, P〈0.001）. Conclusion： PAR1 play an important role in thrombin-mediated lung cancer cell function responses, indicating that PAR1 can be seen as an attractive therapeutic target in lung cancer therapy.
出处
《生物技术通讯》
CAS
2012年第4期497-502,共6页
Letters in Biotechnology
关键词
蛋白酶活化受体1
凝血酶
肺癌
迁移
黏附
克隆形成
胶原收缩
protease-activated receptor 1
thrombin
lung cancer
migration
adhesion
proliferation
collagen contraction
