摘要
目的研究siRNA-ΔNp63干扰质粒在体外对宫颈癌HeLa细胞迁移、侵袭能力的影响。方法在体外利用siRNA-ΔNp63干扰质粒转染HeLa细胞,将实验分为空白对照组、空载体组、阴性质粒组和干扰质粒组;采用Real-time PCR、Western blot检测各实验组细胞中ΔNp63的mRNA、蛋白的表达;通过Transwell侵袭实验、同质黏附实验和异质黏附实验来检测各组细胞的侵袭能力和细胞迁移能力。结果成功将siRNA-ΔNp63干扰质粒转入HeLa细胞;与空白对照组比较,阴性质粒组及空载体组中ΔNp63的mRNA、蛋白的表达水平无明显变化(P>0.05),siRNA-ΔNp63质粒组中ΔNp63的mRNA、蛋白的表达水平表现为降低(P<0.05);空载体组和阴性质粒组平均细胞穿膜数、同质黏附性和异质黏附性无明显变化(P>0.05);而干扰质粒组中平均细胞穿膜数明显降低(P<0.05),同质黏附性增加(P<0.05),异质黏附性明显下降(P<0.05)。结论 siRNA-ΔNp63能明显降低HeLa细胞的迁移和侵袭能力。
Objective To explore the effects of siRNA-ΔNp63 on migration and invasion in HeLa cells in vitro.Methods SiRNA-ΔNp63 was transfected into HeLa cells by lipidosom method in vitro.The cells were divided into control group（without transfection）,empty vector group（only added lipo2000）,negative plasmid group（transfected with negative siRNA） and inferference plasmid group（transfected with effective siRNA）.The mRNA,protein expression level of ΔNp63 were assayed by real-time PCR and Western-blot.The invasion and migration of cell were detected by Transwell,homotypic and hetertypic cell adhesion experiment.Results The siRNA-ΔNp63 interference plasmid into HeLa cells was successful;compared with the control group,mRNA and protein expression of ΔNp63 in negative plasmid group and the empty vector group did not change significantly（P 0.05）.mRNA and protein expression of ΔNp63 in interference plasmid group were decreased（P 0.05）,the average cell penetrating number,homogeneous adhesion and heterogeneity adhesion in negative plasmid group and the empty vector group did not change（P 0.05）,but decreased in the siRNA-ΔNp63 plasmid group about average cell penetrating number（P 0.05）,homogeneous adhesion increase（P 0.05） and heterogeneity adhesion decreased（P 0.05）.Conclusion The siRNA-ΔNp63 can decrease the function of migration and invasion in HeLa cells in vitro.
出处
《中国医药导报》
CAS
2012年第26期20-22,共3页
China Medical Herald
基金
贵州省科学技术基金项目(项目合同编号:黔科合J字[2010]2282号)
