siRNA干扰syk基因对永生化宫颈上皮细胞增殖和侵袭的影响

目的:用小干扰RNA(siRNA)干扰人永生化宫颈鳞状上皮细胞系H8中脾酪氨酸激酶(syk)的表达,观察syk表达降低对细胞增殖和侵袭能力的影响,初步探讨syk在宫颈癌侵袭转移中所起的作用及其表达改变可能影响到的下游靶基因。方法:用siRNA syk干扰永生化宫颈上皮细胞内syk的表达,并以转染control siRNA的细胞作为对照,转染48h后,分别用qRT-PCR和Western blot方法评价干扰效果;在24,48,72h用MTT法检测干扰后细胞增殖情况;Transwell检测细胞侵袭能力;并用qRT-PCR和Western blot方法检测基质金属蛋白酶9(MMP-9)的表达。结果:siRNA干扰syk后能显著降低永生化宫颈上皮细胞系H8内syk mRNA及蛋白的表达(P<0.01)。Syk表达降低并未影响永生化宫颈上皮细胞系H8的增殖(P>0.05),但能使永生化宫颈上皮细胞系H8的侵袭能力显著增强(侵袭细胞数115.57±10.31 vs 39.76±8.79,P<0.01),且syk表达降低后MMP-9的mRNA和蛋白表达均显著增加(P<0.01)。结论:syk表达降低导致永生化宫颈上皮细胞的侵袭能力增加,可能与其表达降低后使MMP-9表达增加相关,syk表达降低与宫颈癌的侵袭转移密切相关。

Effect of syk expression down-regulated by siRNA interference on cell proliferation and invade of human cervical epithelial cells

Objective:To investigate the influence of the expression of spleen tyrosine kinase(syk) down-regulated by siRNA interference on the cell proliferation and invade of human cervical epithelial cells.Methods:The siRNA syk was transfected into human cervical epithelial cells and 48h after the transfection,qRT-PCR and Western blot were used to assay the inhibition effect of siRNA syk on the expression of syk.The cell proliferation of human cervical epithelial cells was determined by MTT.The invasive abilities of human cervical epithelial cells were determined by Transwell.The change of the expression of MMP-9 was detected by qRT-PCR and Western blot.Results:After transfecting with siRNA syk for 48 hours,the expressions of syk mRNA and protein in human cervical epithelial cells were inhibited significantly(P<0.01).After transfecting with siRNA syk,the proliferation of human cervical epithelial cells was not changed(P>0.05).Compared with the control siRNA,the invasive abilities of human cervical epithelial cells were increased significantly(115.57±10.31 vs 39.76±8.79,P<0.01).The expression of MMP-9 was increased significantly(P<0.01).Conclusions:By siRNA interference,the expression of syk gene was effectively inhibited.Silencing syk gene can accelerate the cell invasive abilities by up-regulation of the expression of MMP-9.