miR-219-5p Inhibits Receptor Tyrosine Kinase Pathway by Targeting EGFR in Glioblastoma
Figure 2
Overexpression of miR-219-5p in glioma cell lines decreased proliferation, anchorage independent growth and migration of glioma cells.
A. U138, U343 and U251 cells were transfected with negative control pre-miRs (P-neg) or pre-miRs for miR-219-5p (P-219) and plated in triplicates in 24-well plates. The cell number was counted by using hemocytometer at indicated time points. Average cell number ± SD was plotted. p value was calculated by Students' t test in MS excel. * represents significant p value. For U138, the p values were 0.042, 0.006, 0.003 and 0.003 for Day 2, Day 3, Day 4 and Day 5 respectively; for U343, the p values were 0.06, 0.004, and 0.003 for Day 3, Day 4 and Day 5 respectively; for U251, the p values were 0.009, 0.01 for Day 4 and Day 5 respectively. B. U343 cells were transfected with negative control pre-miRs (P-neg) or pre-miRs for miR-219-5p (P-219) and 300 cells were plated in triplicates in 12-well plates. After 3 weeks of plating, the colonies were stained with crystal violet, photographed (top) and counted (bottom). Average colony number ± SD was plotted. p value was calculated by Students' t test in MS excel. C. U138 and U251 cells were transfected with negative control pre-miRs (P-neg) or pre-miRs for miR-219-5p (P-219). After 36 hrs of transfection, cells were re-plated on the soft agar plates in triplicates. After 3 weeks, the soft agar colonies were counted under microscope. Average colony number ± SD was plotted. p value was calculated by Students' t test in MS excel D. U138 cells were transfected with negative control pre-miRs (P-neg) or pre-miRs for miR-219-5p (P-219). After 72 hrs of transfection, cells were counted and plated for matrigel migration assay and after 18 hrs, cells were fixed, stained with crystal violet, photographed. E. Quantification of D. Average cell number ± SD was plotted. p value was calculated by Students' t test in MS excel.