RSV Effect on Cancer Cell Growth

To measure the effect of the RSV on the growth of HCC and colon cancer cells, an MTT assay was performed with a 3-(4,5 dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma-Aldrich, ST. Louis, MO, USA) solution as described previously (29 (link), 30) (link). The cells were seeded in 48-well plates at a density of 1 × 10⁴ cells/well (BNL-HCC, Hep3B, CT-26, HCT-116 and LoVo) or 5 × 10³ cells/well (Huh-7, SNU-739, HT-29). After 24 h cell culture, the cells were infected with RSV at a MOI of 0.1 and incubated at 37℃ in a 5% CO₂ incubator for three and five days. After incubation, the supernatant was removed and 100 μl of MTT solution (2 mg/ml MTT in autoclaved distilled water) was added to each well. The plates were incubated for 4 h at 37℃, and the culture media were removed. Then 100 μl of DMSO was added for solubilization of the formazan crystals. The color development was analyzed at 595 nm by a Spectra Max 250 microplate reader (Molecular Devices, Sunnyvale, CA, USA).