RNAi干扰电压门控钠通道scn8a基因对人宫颈癌SiHa细胞侵袭转移的影响

目的研究scn8a基因(编码Nav1.6亚型钠离子通道蛋白)对人宫颈癌SiHa细胞增生、迁移和侵袭能力的影响。方法采用小干扰RNA技术(small interfering RNA,siRNA)下调人宫颈癌SiHa细胞scn8a基因表达,使用半定量RT-PCR和Westernblotting方法分别检测siRNA转染细胞后scn8a mRNA和Nav1.6蛋白变化,噻唑蓝比色法检测其对SiHa细胞的增生影响,采用划痕实验、Matrigel侵袭实验观察其对细胞迁移和侵袭能力的影响。结果 siRNA转染SiHa细胞48 h后,scn8a mRNA和Nav1.6蛋白水平分别降低62.7%(P<0.05)和65.8%(P<0.05),针对scn8a基因的siRNA对SiHa细胞的增生无影响(P>0.05),但可有效抑制细胞迁移达55.6%(P<0.05)和侵袭达36.9%(P<0.05)。结论针对scn8a的siRNA干扰了电压门控钠通道在宫颈癌SiHa细胞中表达,其对细胞增生无影响,但抑制了细胞的迁移和侵袭能力。

Effect of RNA interference targeting scn8a gene on migration and invasion of cervical cancer SiHa cells

Objective To study the effects of voltage-gated sodium channel scn8a gene (Nav1.6 subtype) on proliferation, migration and invasion of human cervical cancer SiHa cells. Methods After transfection with small interfering RNA (siRNA) targeting scn8a ,the expressions of scn8a mRNA and Nav1.6 protein in human cervical cancer SiHa cells were determined by RT-PCR and Western blotting, respectively. MTT method was used to examine the effect of scn8a expression on proliferation of the cells. "wound healing" and Matrigel invasion test were used to detect the effect of scn8a expression on migration and invasion of the cells, respectively. Results After 48 h of transfection with siRNA targeting scn8a, the expression of relative scn8a mRNA and Nav1.6 protein levels were decreased by 62.7% (P<0.05) and 65.8% (P<0.05), respectively. The proliferation of SiHa cells at 48 h after siRNA targeting scn8a was shown no changes. The migration of SiHa cells measured by "wound healing"after siRNA targeting scn8a was inhibited by 55.6% (P<0.05), whilst Matrigel invasion of the cells was significantly reduced by 36.9% after siRNA transfection targeting scn8a (P<0.05). Conclusion siRNA targeting scn8a gene had no effect on proliferation of cervical cancer SiHa cells, but suppressed both migration and invasion of the cells.