siRNA沉默S100A4基因表达抑制甲状腺癌细胞侵袭的研究

Study on S100A4 gene silence by siRNA and its inhibition on invasion of human thyroid carcinoma cell

目的 探讨S100A4基因对甲状腺癌细胞侵袭的影响和可能机制.方法 采用S100A4基因小干扰RNA（small interfering RNA,siRNA）转染处理人甲状腺癌ARO细胞后,分别采用荧光实时定量RT-PCR和Western blot检测S100A4基因和基质金属蛋白酶-2mRNA和蛋白水平;分别采用软琼脂集落培养试验和Boyden小室模型试验检测癌细胞的锚着不依赖性增殖和侵袭能力.结果 siRNA转染组S100A4基因mRNA和蛋白水平明显下调,且呈浓度依赖性（P＜0.000 1）.siRNA转染组软琼脂集落形成数和穿过滤膜的细胞数均明显下降,且呈浓度依赖性（P＜0.005;P＜0.005）.S100A4转染组MMP-2基因mRNA和蛋白水平明显下调.结论 采用S100A4 siRNA转染可抑制甲状腺癌细胞侵袭转移,其机制可能与下调MMP-2表达有关.

Objective To explore the effects and mechanism of S100A4 gene silence on invasion of human thyroid cancer cell. Methods After thyroid cancer cell ARO was transfected by S100A4 small interfering RNA （siRNA）, mRNA and protein level of S100A4 and matrix metalloproteinase 2 （MMP-2） were determined by real time RT-PCR and Western blot respectively. The anchorage-independent growth was examined by colony formation assay in soft agar, and invasion ability was evaluated by boyden chamber model. Results The level of mRNA and protein of S100A4 was significantly inhibited in ARO cancer cells transfected by S100A4 siRNA.Transfection with S100A4 siRNA could inhibit anchorage-independent growth and invasion ability of thyroid cancer cell ARO in a dose-dependent manner. mRNA and protein expression of MMP-2 were down-regulated by S100A4 siRNA. Conclusion S100A4 siRNA can inhibit the invasion of thyroid cancer cell through down-regulation of MMP-2.