Enhancement of survivin gene downregulation and cell apoptosis by a novel combination: liposome microbubbles and ultrasound exposure.
Ultrasound-mediated microbubble destruction (sonoporation) is an efficient and safe nonviral technique for gene delivery. In the present work, we hypothesized that short hairpin RNA (shRNA) interference therapy targeting human Survivin gene could be transfected by the novel combination of ultrasound exposure (USE) and liposome microbubbles (LM). ShRNA vectors targeting Survivin were constructed and transfected under USE and LM conditions. The optimal transfection efficiency and cell injury were compared with those of polyethylenimine (PEI)-mediated transfection in different cancer cell lines (HeLa, HepG2, Ishikawa, MCF-7, and B16-F10). The effects of gene downregulation and cell apoptosis were further investigated. The results indicated that P + USE + LM group could significantly increase the gene expression as compared with plasmid group, plasmid + USE group, plasmid + LM group (P < 0.001). The transfection efficiency of the novel combination was nearly equal to PEI-mediated transfection in some cancer cell lines while the cell viability did not decrease markedly. Reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis also confirmed that Survivin mRNA and protein expression could be knocked down significantly by shRNA transfection under USE and LM condition (P < 0.001). This is the first study to verify the role of shRNA therapy in vitro with novel combination of USE and LM. We concluded that this nonviral technique would be valuable in the gene transfection of shRNA and Survivin gene downregulation would lead to apparent cell apoptosis.
Chen
Zhiyi
Z
Department of Medical Ultrasound, The Third Affiliated Hospital of Guangzhou Medical College, Guangzhou, 510150, China. winchen1408@126.com
Liang
Kun
K
Liu
Jianhua
J
Xie
Mingxing
M
Wang
Xinfang
X
Lü
Qing
Q
Zhang
Jing
J
Fang
Lingyun
L
D017209:Apoptosis; D003131:Combined Modality Therapy; D015536:Down-Regulation; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000627:therapeutic use; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008081:Liposomes; D045423:Microbubbles; D008869:Microtubule-Associated Proteins / Q000235:genetics* / Q000378:metabolism; D009369:Neoplasms / Q000235:genetics / Q000473:pathology* / Q000628:therapy*; D034741:RNA, Small Interfering / Q000494:pharmacology*; D000077022:Survivin; D014162:Transfection; D014407:Tumor Cells, Cultured; D014465:Ultrasonics
C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008081:Liposomes; D008869:Microtubule-Associated Proteins; D034741:RNA, Small Interfering; D000077022:Survivin