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. 2022 Jul 4;18(11):4341–4356. doi: 10.7150/ijbs.71134

Figure 2.

Figure 2

Activation of VIPR1 by VIP inhibits HCC cell growth and migration in vitro. (A) Ki-67 immunofluorescence staining of Control (Ctrl) and VIP (200nM) treated HCC cell lines (HepG2, Hep3B and Huh-7) (scale bars, 200μm). (B) Representative images of colony formation in Ctrl and VIP treated groups (Hep3B and Huh-7 cells were seeded at 1000 cells per well; HepG2 cell were seeded at 5000 cells per well. The cells were kept in culture for15 days). (C) Cell proliferative capacity was monitored by CCK-8 assay for about 72h. (D) Western Blot analysis of cyclin E and cyclin D1 in Ctrl and VIP treated cells. (E) Phalloidin staining for cytoskeletal structure in Vector-HCCLM3 cell and Vector-MHCC97H cell (shown as Vector) and VIP treated lenti-VIPR1 overexpressing HCCLM3 cell and MHCC97H cell (namely the VIP/VIPR1 signaling activation group, shown as VIP+VIPR1). Blue: DAPI; Red: F-Actin. (F) Wound healing assay of placebo (PBS) treated Vector-HCCLM3 (shown as Vector); VIP treated lenti-VIPR1 overexpressing HCCLM3 cell (shown as VIPR1+VIP), VIP treated Vector-HCCLM3 cell (shown as Vector+VIP). All groups were treated for 24h in vitro. Two-paired unpaired Student's t test was performed in (A). Values represent means±SEM. *P< 0.05, **P< 0.01, ***P< 0.001.