Figure 1
Knockdown of ASncmtRNA reduces viability and tumorigenic potential of human breast cancer cells. (A) MDA-MB-231, MCF7 and ZR-75-1 human breast cancer cells were transfected for 24 h with 200 nM ASO-1537S or ASO-C, or with transfection agent alone and cell death was measured by Trypan Blue (Tb) exclusion assay. (B,C) Death of MDA-MB-231 cells treated as in (A) for 24 and 48 h was determined by Tb (B) and propidium iodide (PI) (C) exclusion assays. (D) Live cells/well were evaluated by Tb exclusion after 24, 48 and 72 h. (E) MDA-MB-231 cells treated as in (A) were cultured in Matrigel-coated Boyden chamber inserts for 48 h. Inserts were fixed, stained with DAPI and nuclei were counted. (F) Anchorage-independent growth was evaluated in 12-well plates, in which 2 × 103 Tb-negative MDA-MB-231 cells, transfected as in (A), were seeded onto soft agar. Colony formation capacity was evaluated after 21 days in culture. (G) Whole-well microphotographs of colonies and zoom-in under phase contrast microscopy at 4X and 10X magnification. All quantitative data shows average measurement from three independent experiments in triplicate. Statistically significant differences with respect to non-treated cells are indicated (**p < 0.01, ***p < 0.001). L, cells treated with Lipofectamine 2000; C, cells transfected with ASO-C and 1537 S, cells transfected with ASO-1537S.
