RNA干扰沉默Fra-1基因对膀胱癌T24细胞侵袭和迁移的影响

Effects of Fra-1 gene silencing by small interfering RNA on invasion and metastasis of bladder cancer T24 cells

目的研究RNA干扰沉默Fra-1基因表达对人膀胱癌T24细胞侵袭及迁移能力的影响。方法设计合成Fra-1特异性si RNA并转染人膀胱癌T24细胞。实验设空白对照组(转染时只加脂质体)、阴性对照组(无关序列si RNA转染)和干扰组(Fra-1特异si RNA转染)。采用RT-PCR检测Fra-1、MMP-2及MMP-9m RNA水平。Western blot检测Fra-1、MMP-2、MMP-9、STAT3及磷酸化STAT3(p-STAT3)蛋白表达水平,Transwell小室法和划痕实验检测细胞侵袭和迁移能力。结果干扰组T24细胞Fra-1、MMP-2及MMP-9m RNA水平较其他两组明显降低(P<0.01),并且其MMP-2、MMP-9及p-STAT3蛋白表达受到明显抑制(P<0.01)。干扰组T24细胞侵袭力和迁移力较其他两组明显下降(P<0.01)。结论 RNA干扰可有效抑制T24细胞中Fra-1基因的表达,降低细胞侵袭及迁移能力,其机制可能与STAT3/MMPs信号通路有关。

[Objective] To investigate the effects of Fra-1 gene silencing by small interfering RNA （siRNA） on invasion and migration of bladder cancer T24 cells in vitro. [Methods] The Fra-1 siRNA was constructed and then transfected into T24 cells in vitro. The blank control group, negative control group and siRNA transfection group were designed in this study. The expression level of Fra-1, MMP-2 and MMP-9 mRNAs were detected by RT-PCR. The expressions of Fra-1, MMP-2, MMP-9, total STAT3 and p-STAT3 proteins were detected by Western blot. The invasion and migration capabilities were evaluated by using Transwell chamber migration assay and scratch wound healing assay, respectively. [Results] The levels of Fra-1 mRNA, MMP-2 and MMP-9 mRNAs in the siRNA transfection group were significantly decreased compared with that in other groups （P〈0.01）, so were the expressions of MMP-2, MMP-9 and p-STAT3 proteins （P〈 0.01）. In the siRNA transfection group, the invasion and migration capabilities of T24 cells were also signifi- candy decreased compared with those in other groups （P〈 0.01）. [Conclusions] Small interfering RNA can ef- ficiently inhibit the expression of Fra-1, which results in decreased capacities of invasion and migration of T24 cells. The mechanism may be involved in STAT3/MMPs signaling pathway.