Figure 7.
The contribution of HMGB1 on the effect of circ_0007385 knockdown in NSCLC cells in vitro. (a) Western blotting measured HMGB1 protein level in A549 and H1975 cells transfected with HMGB1 overexpression vector (pEGFP‐C1‐HMGB1, HMGB1) or its control vector (vector) () vector, () HMGB1. (b–j) A549 and H1975 cells were transfected with sh‐circ or sh‐NC, and cotransfected with sh‐circ and either HMGB1 or vector. (b and c) CCK‐8 assessed OD450 nm value on 24, 48 and 72 hours (
) sh‐NC, (
) sh‐circ, (
) sh‐circ+vector, (
) sh‐circ+HMGB1; () sh‐NC, () sh‐circ, () sh‐circ+vector, () sh‐circ+HMGB1. (d and e) Transwell assay determined the numbers of migratory and invasive cells (
) sh‐NC, (
) sh‐circ, (
) sh‐circ+vector, (
) sh‐circ+HMGB1; () sh‐NC, () sh‐circ, () sh‐circ+vector, () sh‐circ+HMGB1. (f) Western blotting detected protein expression of the PCNA, E‐cad and N‐cad () sh‐NC, () sh‐circ, () sh‐circ+vector, () sh‐circ+HMGB1; () sh‐NC, () sh‐circ, () sh‐circ+vector, () sh‐circ+HMGB1. (g–j) Transfected A549 and H1975 cells were treated with DDP for 48 hours. (g) CCK‐8 measured IC50 of DDP after treatment of 0, 2.5, 5, 10, 20, 40, and 80 μM of DDP () sh‐NC, () sh‐circ, () sh‐circ+vector, () sh‐circ+HMGB1. (h) FCM examined apoptosis rate (%) () sh‐NC, () sh‐circ, () sh‐circ+vector, () sh‐circ+HMGB1 and (i and j) caspase‐3 activity after treatment of 10 μM of DDP () sh‐NC, () sh‐circ, () sh‐circ+vector, () sh‐circ+HMGB1; () sh‐NC, () sh‐circ, () sh‐circ+vector, () sh‐circ+HMGB1. **P < 0.01.