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目的通过RNA干扰(RNA interference,RNAi)抑制Hep-2细胞中水通道蛋白-1(aquaporin-1,AQP1)的表达,探讨AQP1对喉癌细胞黏附、运动及侵袭能力的影响。方法构建AQP1特异性的siRNA重组质粒,免疫组化检测RNAi抑制AQP1表达的效果;分别用细胞基质黏附实验、Transwell小室和Matrigel基质侵袭实验检测细胞黏附、运动和侵袭能力。结果AQP1-siRNA重组质粒有效的抑制了Hep-2细胞中AQP1的表达。细胞基质黏附实验结果显示:Hep-2细胞与细胞外基质的黏附率,AQP1-siRNA实验组为(41.6+4.2)%与阴性对照组(43.6+3.9)%和空白对照组(45.2±4.0)%无明显差异(P均>0.05);体外细胞运动和侵袭实验结果显示:Hep-2的穿膜细胞数,实验组分别为(36±3)和(23±4)个/高倍镜,显著低于空白对照组的(70±5)和(65±4)个/高倍镜以及阴性对照组的(72±4)和(69±4)个/高倍镜(P均<0.01),而后两组细胞间运动和侵袭能力方面差异均无显著性(P>0.05)。结论AQP1RNAi重组质粒能够阻断AQP1的表达,对Hep-2细胞的黏附力没有影响,但能够抑制喉癌细胞的运动和侵袭能力,与喉癌的转移机制密切相关。
Objective To investigate the effect of AQP1 on adhesion, motility and invasion of laryngeal carcinoma cells by inhibiting the expression of aquaporin-1 (AQP1) in Hep-2 cells by RNA interference (RNAi). Methods AQP1 specific siRNA recombinant plasmids were constructed. The effect of RNAi on AQP1 expression was detected by immunohistochemistry. Cell adhesion, motility and invasion were evaluated by cell adhesion assay, Transwell chamber and Matrigel invasion assay. Results AQP1-siRNA recombinant plasmid effectively inhibited AQP1 expression in Hep-2 cells. The adhesion rate of Hep-2 cells to extracellular matrix was (41.6 ± 4.2)% in the AQP1-siRNA group and (43.6 ± 3.9)% in the AQP1-siRNA group and 45.2 ± 4.0 in the blank control group % (P> 0.05). The results of in vitro cell motility and invasion showed that the number of transmembrane cells in Hep-2 cells was (36 ± 3) and (23 ± 4) / (70 ± 5) and (65 ± 4) / high power of the control group and (72 ± 4) and (69 ± 4) / high power of the negative control group (P <0.01) There were no significant differences in cell motility and invasiveness between groups (P> 0.05). Conclusion AQP1RNAi recombinant plasmid can block the expression of AQP1 and has no effect on the adhesion of Hep-2 cells, but it can inhibit the movement and invasion ability of laryngeal carcinoma cells, which is closely related to the metastasis mechanism of laryngeal carcinoma.