Fig. 3: H19 mediates E2-induced stem-like properties in PTC cells.

a TPC-1 and K-1 cells were treated with various concentrations (0, 10, and 50 nM) of E2 for 36 h. Total RNA was extracted and subjected to detect H19 expression by RT-qPCR analysis. The relative H19 level was normalized to ACTB. Data were shown as means ± SD. (n = 3, **P < 0.01 and ***P < 0.001). b TPC-1 and K-1 cells were treated with 50 nM E2 for 0, 4, 8, 12, 24, 36 h. Total RNA was extracted and subjected to detect H19 expression by RT-qPCR analysis. The relative H19 level was normalized to ACTB. Data were shown as means ± SD. (n = 3, *P < 0.05). c TPC-1 and K-1 cells were treated with E2 (50 nM) for 36 h. Total RNA was extracted and subjected to detect pre-H19 expression by RT-qPCR. The relative pre-H19 levels were normalized to ACTB. Data were shown as means ± SD. (n = 3, **P < 0.01 and ***P < 0.001). d Total RNA was extracted in NTC, shERβ-1, shERβ-2 K-1 cells. Pre-H19 and H19 were detected by RT-qPCR assay. The relative RNA levels were normalized to ACTB. Data were shown as means ± SD (n = 3, **P < 0.01 and ***P < 0.001). e pGL3-EV, pGL3-H19-WT, -H19-Del, or -H19-Mut along with pRL-SV40 were transfected into K-1 cells with E2 treatment. After 24 h, dual-luciferase reporter assays were performed (n = 3, *P < 0.05). f siNC and siERβ were co-transfected into K-1 cells with pGL3-EV, pGL3-H19-WT, -H19-Del, or -H19-Mut along with pRL-SV40. After 24 h, dual-luciferase reporter assays were performed (n = 3, ***P < 0.001). g Sphere formation abilities of K-1 under different conditions were compared. Representative images were presented, the scale bar represents 100 μm. The numbers and size of spheres were counted after culture for 10 days. Data were shown as means ± SD. (n = 3, *P < 0.05, **P < 0.01 and ***P < 0.001)