Effective sequence screening of siRNAs targeted protein kinase C ? and their effects on A549 cell line / 基础医学与临床

ABSTRACT

Objective To investigate the effects of siRNAs targeted protein kinase C ?(PKC?) on the proliferation and apoptosis of A549 cell line.Methods Six PKC? siRNAs were designed and chemical synthesized. Candidate PKC? siRNA was transfected into A549 cells,PKC? mRNA level was determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Then,the effects of more effective PKC? siRNAs on A549 were further investigated by using clone formation assay,Hoechst 33258 staining,propidium iodide (PI) staining,Annexin V-FITC and PI dual staining and western blot. Results Six candidate PKC? siRNAs were designed and named as No.1,No.2,No.3,No.4,No.5 and No.6 PKC? siRNA. Compared with controls,PKC? mRNA levels were all significantly downregulated and the cell proliferation was inhibited in A549 cells treated with candidate PKC? siRNAs except No.5 (P