(A) HCT-116 cells were treated with DMSO, 0.005, 0.05, or
0.5 μM of 1 and incubated for 24 hr. Cells were then imaged,
and viability was determined using the XTT cell-viability assay. Cell
proliferation was inhibited at 0.05 μM and cell death was observed at 0.5
μM. ns = not significant, *p < 0.01, ** p < 0.001.
(B) HCT-116 cells were treated with 50 nM 1 and
total RNA was extracted from various time points (0–24 hr) for RT-qPCR
analysis. Expression levels of miRs −21, −1246, −203a-3p,
−200b-3p, and −361 and pre-miR-21 were analyzed using U6 snRNA as
reference gene. miR-1246 did not show significant amplification up to 24 hr.
(C) To show if miR-21 can rescue bPGN treated cells, HCT-116
cells were transfected with miR-21 mimic 4–5 hr before the cells were
treated with 100 nM bPGN (2× GI50). Cell
viability was measured at 24 and 48 hr by XTT assay. *p < 0.05. All
experiments were performed in triplicate.