RNA 干扰 HDAC1对人宫颈癌SiHa细胞迁移及侵袭影响

目的 观察应用RNA干扰技术沉默组蛋白去乙酰化酶1（HDAC1）对人宫颈癌SiHa细胞迁移和侵袭的影响，并探讨其可能机制。方法 体外合成针对HDAC1的小干扰RNA（siRNA），采用脂质体法转染人宫颈癌SiHa细胞。采用Western blot法检测细胞HDAC1蛋白和乙酰化组蛋白H4的表达，荧光定量PCR方法检测HDAC1、核因子-κB （NF-κB）和尿激酶型纤溶酶原激活剂（uPA）基因mRNA表达，划痕实验检测细胞迁移能力，Transwell侵袭实验检测细胞侵袭能力。结果 经转染HDAC1基因siRNA后，宫颈癌SiHa细胞HDAC1 mRNA和蛋白表达降低，NF-κB和uPA基因mRNA表达也降低，细胞内组蛋白H4乙酰化水平增加。SiHa细胞迁移速度降低，穿过Transwell滤膜的细胞数量减少。结论 HDAC1基因siRNA能够通过抑制宫颈癌SiHa细胞HDAC1基因表达，抑制人宫颈癌SiHa细胞迁移和侵袭能力。提高组蛋白乙酰化水平，下调NF-κB和uPA基因表达可能是其作用机制。

EFFECT OF RNA INTERFERENCE TARGETING HDACl GENE ON MIGRATION AND INVASION OF CELL LINE SIHA IN HU- MAN CERVICAL CARCINOMA

Objective To observe histone deacetylase 1 （HDAC1） gene silencing on migration and invasion in human cervical carcinoma cell line SiHa, and investigate its possible mechanism. Methods The small interference RNA （siRNA） targeting HDAC1 was generated in vitro, and transfected into human cervical carcinoma SiHa cells by Lipofectamine 2000. Western blot was used to detect the HDAC1 protein and acetyl level of histone H4, and real time PCR was used to detect mRNA of HDAC1, NF-κB, and uPA genes. Cell migration was estimated by wound-healing assay and cell invasion estimated by Transwell assay. Results After siRNA targeting HDAC1 had been transfected into SiHa cells, the expressions of HDAC1 gene protein and mRNA decreased, and the acetyl level of histone H4 in SiHa cells increased. The cell migration and invasion activity declined. Meanwhile, the mRNA expressions of NF-κB and uPA gene in SiHa cells decreased. Conclusion The siRNA targeting HDAC1 can inhibit the cellular migration and invasion activity in cervical cancer cells through suppressing the expression of HDAC1. Increasing acetyl level of histone to down-regulate NF-κB and uPA gene expressions may be its mechanism.