Figure 7.

miR-101 regulates multiple cell-cycle-related genes and controls the growth of HCC cells. (A) Distribution of objects from the miR-101-regulated sub-network in the KEGG Cell-cycle pathway. The genes labeled with grey color indicate the nodes in miR-101-regulated subnetwork. (B–C) Expression levels of genes that are the first-layered regulators (B) and second-layered effectors, which are cell-cycle-related genes (C) of the cell cycle in HepG2 cells transfected with negative control and miR-101. Total RNA was obtained 48 h after transfection, and mRNA measurements were performed in duplicate. Data are presented as the means ± SEM of three independent experiments. (D) Effect of miR-101 on growth of HepG2 cells. HepG2 cells were transfected with control or miR-101 and maintained for 2 and 4 days, respectively. The cells were fixed and stained with DAPI. The cell number was determined using an InCell 1000 imaging system. Data are the average of nine fields. (E) Effects of miR-101 on colony-forming activity of cultured HepG2 cells. Cells were transfected with control or miR-101 and plated at 2000 cells/well in six-well plates for 14 days. After staining with crystal violet, the colony number and size were determined using Image J.