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. 2021 Mar 9;11:638171. doi: 10.3389/fonc.2021.638171

Figure 4.

Figure 4

GPER-mediated EGFR/ERK signaling decreases cell viability in HCC cell lines. The GPER-specific agonist G1 blocked progression to the S phase of the cell cycle (A) and promoted apoptosis (B) in HCCLM3 cells (P < 0.05; a vs. ctrl). Synchronized cells were cultured with G1 and stained with propidium iodide or Annexin V-FITC/PI, and the cell cycle distribution and apoptosis were measured using flow cytometry. (C, D) The observed cell cycle and apoptosis blockade by G1 were reversed by GPER interference (P < 0.05; a vs. ctrl + siRNA, b vs. G1 + siGPER). (E, F) The observed cell cycle and apoptosis blockade by G1 were attenuated by specific inhibitors targeting EGFR (AG) or ERK (U0126), but not AKT (WM) (P < 0.05; a vs. ctrl, b vs. G1 + AG, c vs. G1 + U0126). The above data represents the mean ± SD from triplicate independent experiments. (G) The G1-inhibited cell growth was reversed by GPER interference (P < 0.05; a vs. ctrl + siRNA, b vs. G1 + siGPER). (H) The observed inhibition of cell growth by G1 was attenuated by AG and U0126 (P < 0.05; a vs. ctrl, b vs. G1 + AG, c vs. G1 + U0126). The above data represents the mean ± SD from five different experiments.