siRNA-ILK慢病毒载体的构建

目的构建RNA干扰整合素连接激酶(ILK)基因的慢病毒载体并转染胰腺癌细胞(Panc-1),并验证其干扰有效性。方法对胰腺癌Panc-1细胞进行细胞培养,成功构建ILK-specific sh RNA慢病毒载体后对Panc-1细胞进行转染,荧光显微镜下观察,评估转染效率,然后通过Real time-PCR及Western Blot方法验证干扰基因片段的有效性。结果成功构建si RNA-ILK慢病毒载体并转染Panc-1细胞,荧光显微镜下观察可见转染效率达80%以上,RNAi靶点病毒转染的细胞组(KD)的ILK m RNA及蛋白表达明显低于空蛋白组(NC)及正常细胞组(CON,0.01)。结论成功构建RNA干扰ILK基因的慢病毒载体并转染胰腺癌细胞(Panc-1),验证了RNA干扰片段的有效性。

Construction of siRNA-ILK lentivirus vectors

Objective To construct and transfect si RNA-ILK gene lentivirus vector into the pancreatic cancer cells（Panc-1） to verify the interference effectiveness. Methods ILK-specific sh RNA lentivirus vector was constructed and transfected into Panc-1 cells which were observed by fluorescence microscope, and Real time-PCR and Western blot were used respectively to verify the interference effectiveness. Results si RNA-ILK gene lentivirus vector was constructed successfully and transfected into Panc-1 cells, the infection efficiency was more than 80% under fluorescence microscope, the expression level of ILK m RNA and protein was upregulated in transfected group than in negative control or normal control（P〈0.01）. Conclusion The si-ILK gene lentivirus vector was successfully constructed and ILK RNA interference segment was effective.