沉默Pard3基因对宫颈癌细胞系SiHa迁移、侵袭的影响及其机制

Effect of silencing PARD3 gene on migration and invasion of cervical cancer cell line SiHa

目的观察沉默分离缺陷基因3(Pard3)对宫颈癌细胞系SiHa迁移、侵袭的影响,并探讨其机制。方法将SiHa细胞分为siRNA1组、siRNA2组、siRNC组,siRNA1组转染Pard3小干扰RNA1(siRNA1),siRNA2组转染Pard3小干扰RNA2(siRNA2),siRNC组转染空白载体,转染后继续孵育48 h。采用Transwell迁移和侵袭试验观察三组细胞迁移和侵袭能力,分别以细胞迁移相对数量和细胞侵袭相对数量表示。采用实时定量PCR法检测三组细胞中Pard3、JAK2、STAT3、E-cadherin、Vimentin mRNA。采用Western blotting法检测三组细胞中Pard3、JAK2、STAT3、E-cadherin、Vimentin、p-JAK2、p-STAT3蛋白。结果siRNA1组、siRNA2组细胞迁移相对数量和细胞侵袭相对数量均高于siRNC组(P均<0.05)。siRNA1组、siRNA2组细胞中Pard3、E-cadherin mRNA和蛋白相对表达量均低于siRNC组(P均<0.05),siRNA1组、siRNA2组细胞中VimentinmRNA、Vimentin蛋白、p-JAK2蛋白、p-STAT3蛋白相对表达量均高于siRNC组(P均<0.05)。结论沉默Pard3基因可促进SiHa细胞的迁移和侵袭,其机制可能与调节JAK2/STAT3磷酸化信号转导通路有关。

Objective To observe the effect of silencing partitoning defective gene 3(PARD3)on the migration and invasion of cervical cancer cell line SiHa,and to explore its mechanism.Methods SiHa cells were divided into the siR⁃NA1 group,siRNA2 group,and siRNC group.SiHa cells in the siRNA1 group were transfected with PARD3 small inter⁃ference RNA1(siRNA1),the siRNA2 group with PARD3 small interference RNA2(siRNA2),and the siRNC group with blank vector,and then they were incubated for 48 h after transfection.Transwell migration and invasion assay was used to observe the migration and invasion abilities of the three groups,which were represented by the relative number of migration cells and the relative number of invasion cells,respectively.Pard3,JAK2,STAT3,E-cadherin and Vimentin mRNAs were detected by real-time quantitative PCR.Pard3,JAK2,STAT3,E-cadherin,Vimentin,p-JAK2,and p-STAT3 pro⁃teins in the three groups were detected by Western blotting.Results The relative number of migration cells and the rela⁃tive number of invasion cells in the siRNA1 and siRNA2 groups were higher than those in the siRNC group(all P<0.05).The relative expression levels of PARD3,E-cadherin mRNA and protein in the siRNA1 and siRNA2 groups were lower than those in the siRNC group(all P<0.05),and the relative expression levels of Vimentin mRNA,Vimentin protein,p-JAK2 protein and p-STAT3 protein in the siRNA1 and siRNA2 groups were higher than those in the siRNC group(all P<0.05).Conclusion Silencing PARD3 gene can promote the migration and invasion of SiHa cells,and the mechanism may be related to the regulation of JAK2/STAT3 phosphorylation signal transduction pathway.