Research Paper Volume 13, Issue 6 pp 8380—8395

Figure 3. Extracellular HMGB1 increases CD44 expression via upregulating miR-21. (A) Q-PCR analysis shows that rhHMGB1 increases miR-21 expression. HepG2 and HCCLM3 cells were cultured with rhHMGB1 (1μg/ml) for 24h. (B) Immunoblot analysis shows that miR-21 inhibitor represses CD44 expression caused by rhHMGB1. HepG2 and HCCLM3 cells were transfected with negative control or miR-21 inhibitor and then cultured with rhHMGB1 (1μg/ml) for 24h. (C) Q-PCR analysis indicates that miR-21 inhibitor represses CD44 expression caused by rhHMGB1. HepG2 and HCCLM3 cells were transfected with negative control or miR-21 inhibitor and then cultured with rhHMGB1 (1μg/ml) for 24h. (D, E) Results from Invasion experiments indicate miR-21 inhibitor represses HCC invasion caused by rhHMGB1. HepG2 and HCCLM3 cells were transfected with negative control or miR-21 inhibitor and then cultured with rhHMGB1 (1μg/ml) for 24h. (F–H) Results from sphere formations experiments indicate miR-21 inhibitor represses HCC sphere formations caused by rhHMGB1. HepG2 and HCCLM3 cells were transfected with negative control or miR-21 inhibitor and then cultured with rhHMGB1 (1μg/ml) for 24h. (G) Results from metastasis model in vivo show that miR-21 inhibitor suppresses liver metastasis caused by rhHMGB1. HepG2 cells were transfected with negative control or miR-21 inhibitor and then cultured with rhHMGB1 (1μg/ml) for 24h. Data are means ± SEM, * means p<0.05, ** means p<0.01, *** means p<0.001 by unpaired student T test.