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目的探讨华支睾吸虫重组14-3-3ε蛋白(r Cs14-3-3ε)对肝内胆管癌细胞株CCLP-1细胞波形蛋白(vimentin)表达的影响,观察该过程中CCLP-1细胞侵袭和迁移能力的变化。方法用不同浓度(1、5、10μg/ml)r Cs14-3-3ε与CCLP-1细胞共孵育24、48、72 h后,通过Q-PCR、Western blot等实验方法检测vimentin的表达水平。用划痕愈合实验和Transwell小室侵袭实验检测r Cs14-3-3ε对CCLP-1细胞迁移和侵袭能力的影响。结果 Q-PCR实验表明5μg/ml r Cs14-3-3ε刺激CCLP-1细胞72 h后及10μg/ml r Cs14-3-3ε刺激CCLP-1细胞24、48、72 h后,vimentin m RNA水平显著上调;Western blot实验显示,10μg/ml r Cs14-3-3ε刺激CCLP1细胞48、72 h后,vimentin蛋白水平显著上调。划痕实验表明,r Cs14-3-3ε(10μg/ml)能够促进细胞的迁移能力,处理后的细胞迁移率增大(P<0.01);Transwell小室检测显示,r Cs14-3-3ε(10μg/ml)处理组中穿膜细胞数为对照组的1.78倍,显著增加(P<0.05)。结论 r Cs14-3-3ε可以通过上调vimentin蛋白的表达,从而促进肝内胆管癌细胞株CCLP-1的侵袭和迁移能力。
Objective To investigate the effect of Cs14-3-3ε on the expression of vimentin in human hepatocellular carcinoma cell line CCLP-1 and to investigate the effect of Cs14-3-3ε on the invasion of CCLP-1 cells And changes in the ability to migrate. Methods After co-cultured with CCLP-1 and Cs14-3-3ε at different concentrations (1,5, 10μg / ml) for 24,48,72 h, the expression of vimentin was detected by Q-PCR and Western blot. Scratch healing assay and Transwell chamber invasion assay were used to determine the effect of Cs14-3-3ε on the migration and invasion of CCLP-1 cells. Results The results of Q-PCR showed that after stimulating CCLP-1 cells with 5μg / ml Cs14-3-3ε for 72 hours and stimulating CCLP-1 cells with Cs14-3-3ε 10μg / ml for 24,48,72 h, the level of vimentin m RNA Western blot analysis showed that vimentin protein level was significantly up-regulated in CsCl-3 cells stimulated with 10μg / ml CsCl-4 for 48 and 72 h. Scratch assay showed that Cs14-3-3ε (10μg / ml) promoted cell migration and the cell migration rate increased (P <0.01). Transwell assay showed that Cs14-3-3ε (10μg / ml) / ml), the number of transmembrane cells in the treatment group was 1.78 times that of the control group, which was significantly increased (P <0.05). Conclusion Cs14-3-3ε can promote the invasion and migration of CCLP-1 cells by up-regulating the expression of vimentin protein.