Exosomal miR-182 regulates the effect of RECK on gallbladder cancer

Figure 1 Western blot analysis of cluster of differentiation 63 and cluster of differentiation 81 exosomes. A: Expression levels of cluster of differentiation (CD) 63 and CD81; B: Western blot bands. Exosomes were isolated from cells; the isolation process is described in “Separation and screening of exosomes.”

Figure 2 MicroRNA 182 was highly expressed, whereas there was low expression of reversion-inducing-cysteine-rich protein with kazal motifs in gallbladder cancer. A: MicroRNA 182 (miR-182) was highly expressed in gallbladder cancer (GC) tissues; B: Exosomal miR-182 was highly expressed in patients with GC; C: MiR-182 was highly expressed in GC cells; D: Reversion-inducing-cysteine-rich protein with kazal motifs (RECK) protein was highly expressed in GC tissues; E: RECK protein was highly expressed in GC cells. ^aP < 0.05, ^bP < 0.01 vs HGBEC. ^cP < 0.001.

Figure 3 Relationship between microRNA 182 and tumor-node-metastasis staging. A: MicroRNA 192 (miR-182) was highly expressed in the N1/N2/N3 phase; B: Exosomal miR-182 was highly expressed in the N1/N2/N3 phase; C: MiR-182 was highly expressed in the M1 phase; D: Exosomal miR-182 was highly expressed in the M1 phase. ^cP < 0.001.

Figure 4 Exosomal microRNA 182 can promote cell proliferation, migration and invasion, and inhibit apoptosis and expression of reversion-inducing-cysteine-rich protein with kazal motifs. A: Exosomal microRNA 182 (miR-182) enhanced the viability of gallbladder cancer (GC) cells; B: Exosomal miR-182 promoted cell invasion and migration; C: Effects of exosomal miR-182 on caspase 3, caspase 9, E-cadherin, B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein, N-cadherin, and β-catenin in GC cells; D: Exosomal miR-182 inhibited apoptosis; E: Exosomal miR-182 inhibited reversion-inducing-cysteine-rich protein with kazal motifs expression. ^aP < 0.05, ^bP < 0.01, and ^cP < 0.001 vs the negative control (NC) group.

Figure 5 Reversion-inducing-cysteine-rich protein with kazal motifs inhibited cell proliferation, migration and invasion, and promoted apoptosis. A: Reversion-inducing-cysteine-rich protein with kazal motifs (RECK) inhibited viability of gallbladder cancer (GC) cells; B: RECK inhibited cell invasion and migration; C: Effects of exosomal microRNA 182 on caspase 3, caspase 9, E-cadherin, B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein N-cadherin, and β-catenin in GC cells; D: RECK promoted apoptosis; ^aP < 0.05, and ^bP < 0.01 vs the negative control (NC) group.

Figure 6 MiR-182 targeted inhibition of reversion-inducing-cysteine-rich protein with kazal motifs expression. A: MicroRNA 182 (miR-182) had binding sites with reversion-inducing-cysteine-rich protein with kazal motifs (RECK) mRNA; B and C: Double luciferase reporter gene assay verified the targeting relationship between miR-182 and RECK; D: MiR-182 was negatively correlated with RECK protein. NC: Negative control.

Figure 7 Rescue experiment. A: Viability comparison of gallbladder cancer cells in each group; B: Comparison of cell invasion in each group; C: Comparison of EH-GB1 cell-related proteins in each group; D: Comparison of apoptosis in each group. ^aP < 0.05 vs the negative control (NC) group. ^cP < 0.05, ^dP < 0.01, and ^fP < 0.001 vs exosomal microRNA 182 group.