抑制Tiam1基因表达促进EC9706细胞侵袭、转移和增强对5-FU化疗敏感性

Tiam1 inhibition promotes invasion and metastasis in EC9706 cells and enhances their chemotherapeutic sensitivity to 5-fluorouracil

目的探讨抑制T淋巴瘤侵袭转移诱导因子1(T lymphoma invasion and metastasis inducing factor 1,Tiam1)基因表达对食管癌EC9706细胞侵袭、转移及对5-FU化疗敏感性的影响。方法靶向Tiam1的3对siRNA转染食管癌EC9706细胞,转染48 h后采用半定量RT-PCR和Western blot法检测各组细胞Tiam1 mRNA和蛋白表达水平的变化。Boyden chamber侵袭小室和划痕实验检测EC9706细胞体外侵袭、转移能力的变化。Tiam1 siRNA与不同浓度5-FU同时作用于EC9706细胞,CCK-8实验检测对EC9706细胞增殖能力的影响;TUNEL法检测对EC9706细胞凋亡能力的影响。结果与对照组相比,转染靶向Tiam1基因的siRNA2和siRNA3能有效抑制EC9706细胞Tiam1 mRNA和蛋白的表达,差异有统计学意义(P<0.05)。Boyden chamber侵袭小室和划痕实验结果显示,与对照组相比,转染Tiam1 siRNA3 48 h后的EC9706细胞侵袭转移能力明显下降,差异有统计学意义(P<0.05)。Tiam1 siRNA3与不同浓度5-FU联合作用均能抑制细胞增殖,与单独5-FU相比差异有统计学意义(P<0.05)。Tiam1 siRNA3与10μg/mL 5-FU联合作用能显著抑制EC9706细胞的凋亡能力与10μg/mL 5-FU单独作用组相比,差异有统计学意义(P<0.05)。结论抑制Tiam1基因表达能有效抑制食管癌细胞的侵袭、转移能力,且能提高EC9706细胞对5-FU的化疗敏感性。

Objective To determine the effects of silencing T lymphoma invasion and metastasis indu- cing factor 1 （ Tiaml ） on the invasion and metastasis capability of esophageal carcinoma cell line EC9706 in vitro and the chemotherapeutic sensitivity of EC9706 to 5-fluorouracil （5-FU）. Methods EC9706 cells were transfected with Tiaml siRNAs. In 48 h after transfeetion, Tiaml mRNA and protein levels were evaluated by RT-PCR and Western blotting. The invasion and metastasis ability of EC9706 cells were checked by Boyden chamber assay and wound healing assay. CCK-8 assay was used to test the inhibitory actions on EC9706 cells after the cells were treated with different concentrations of 5-FU and Tiaml siRNA together. The apoptosis of EC9706 cells was assayed by TUNEL. Results Tiaml siRNA2 and siRNA3 could inhibit the mRNA and protein expression of Tiaml efficiently in EC9706 cells compared with control groups after 48 h （ P 〈 0.05 ）. The results of Boyden chamber assay and wound healing assay showed that the invasion and metastasis ability of EC9706 cells transfeeted with Tiaml siRNA3 for 48 h decreased greatly compared with that in control groups （P 〈 0. 05 ）. The growth inhibitory rates of EC9706 cells treated with different concentrations of 5-FU and Tiaml siRNA3 together were much higher than those treated with different concentrations of 5-FU alone （P 〈 0.05）. The apoptotic rates of EC9706 cells treated with 10 p^g/mL 5-FU and Tiaml siRNA together were also higher than those in control groups （P 〈 0. 05）. Conclusion Inhibiting the expression of Tiaml decreases the invasion and metastasis abilities of EC9706 cells greatly and increases the chemotherapeutic sensitivity of EC9706 cells to 5-FU.