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目的:研究低糖基化E-cadherins对舌鳞癌细胞CAL 27增殖和侵袭的影响,并探讨E-cadherins低糖基化修饰对其介导的细胞间黏附连接(adherens junctions,AJs)稳定性的影响。方法:用编码有FLAG标签的低糖基化Ecadherins和野生型E-cadherins质粒分别转染CAL 27细胞。48 h后,用抗FLAG抗体作为一抗,Western免疫印迹检测外源性E-cadherins的表达;用细胞增殖计数、单层细胞划痕愈合和细胞体外侵袭实验检测细胞的增殖和侵袭能力;免疫沉淀法检测E-cadherins介导的AJs复合体中的α-catenins、β-catenins、γ-catenins和vinculins的含量。采用SPSS17.0软件包对数据进行统计学分析。结果:被转染细胞均有外源性E-cadherins表达。被转染细胞的增殖和侵袭能力比未被转染对照细胞显著减弱(P<0.05),而且低糖基化E-cadherins对CAL 27细胞增殖和侵袭能力的抑制作用比野生型E-cadherins显著增强(P<0.05)。低糖基化E-cadherins介导的AJs复合体中的α-catenins、β-catenins、γ-catenins和vinculins的含量均比野生型E-cadherins介导的AJs复合体显著增多(P<0.01)。结论 :低糖基化Ecadherins比野生型E-cadherins更加显著抑制舌鳞癌细胞的增殖和侵袭,其机制是低糖基化E-cadherins介导的AJs比野生型E-cadherins介导的AJs更加稳定。
Objective: To investigate the effect of hypoglycosylated E-cadherins on the proliferation and invasion of tongue squamous carcinoma cell line CAL 27 and to investigate the effect of E-cadherins hypoglycosylation on the stability of adherens junctions (AJs) . METHODS: CAL27 cells were transfected with FLAG-tagged low glycosylated Ecadherins and wild-type E-cadherins respectively. Forty-eight hours later, anti-FLAG antibody was used as the primary antibody and western blot was used to detect the expression of exogenous E-cadherins. The proliferation and invasion ability of cells were detected by cell proliferation counting, monolayer scratch healing and in vitro cell invasion assay. Precipitation method was used to detect the content of α-catenins, β-catenins, γ-catenins and vinculins in E-cadherins-mediated AJs complex. SPSS17.0 software package for statistical analysis of the data. Results: All transfected cells had exogenous E-cadherins expression. The proliferation and invasion ability of the transfected cells was significantly weaker than that of the untransfected control cells (P <0.05), and the inhibitory effect of low glycosylated E-cadherins on the proliferation and invasion ability of CAL 27 cells was significantly enhanced than that of wild-type E-cadherins (P <0.05). The content of α-catenins, β-catenins, γ-catenins and vinculins in low-glycosylated E-cadherins mediated AJs complex was significantly higher than that of wild-type E-cadherins mediated AJs complex (P <0.01). CONCLUSION: Hypoglycosylated Ecadherins inhibit the proliferation and invasion of tongue squamous carcinoma cells more significantly than wild type E-cadherins. The mechanism is that hypoglycosylated E-cadherins mediated AJs are more stable than wild-type E-cadherins mediated AJs.