Specific subcellular localization of siRNAs delivered by lipoplex in MCF-7 breast cancer cells.
In order to better understand the mechanism of delivery of siRNAs by lipid-based vectors, we investigated the subcellular distribution of siRNAs directed against cyclin D1 delivered by the DLS system in the breast cancer cell line MCF-7. Cells were treated with cyclopentenone or 17beta-estradiol to modulate the level of expression of cyclin D1 mRNA. We qualitatively observed that siRNA localized to specific cytoplasmic compartments in the periphery of the nucleus in granular-like structures that do not correspond to early endosomal vesicles. In cells treated with either cyclopentenone or 17beta-estradiol cellular distribution of siRNAs was not affected but variations in the amount of siRNAs present in cells were found. We suggest these variations might be associated with the effects of cyclopentenone and 17beta-estradiol in cyclin D1 gene expression. Low cytotoxicity and highly cellular uptake of lipoplexes was observed in the presence of serum indicating that the DLS system could be a useful tool for siRNA vectorization in vitro and in vivo.
Lavigne
Carole
C
Laboratoire de biologie cellulaire et de thérapie génique, Département de Biologie, Université de Moncton, 165 Massey Avenue, Moncton, New Brunswick, E1A 3E9, Canada. lavigncar@umoncton.ca
Thierry
Alain R
AR
D001943:Breast Neoplasms / Q000235:genetics / Q000473:pathology; D045744:Cell Line, Tumor; D019938:Cyclin D1 / Q000235:genetics; D003517:Cyclopentanes / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry*; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D034741:RNA, Small Interfering / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000379:methods*; D033921:Vesicular Transport Proteins / Q000378:metabolism
D003517:Cyclopentanes; D008081:Liposomes; D008565:Membrane Proteins; D034741:RNA, Small Interfering; D033921:Vesicular Transport Proteins; C093846:early endosome antigen 1; D019938:Cyclin D1; D004958:Estradiol; C013905:cyclopentenone