Fig. 7.

H19-induced tumour suppression is superior to CAB treatment. a H19 suppresses GH3 tumour growth more potently than CAB treatment (0.75 mg/kg) in vivo. First, 1 × 10⁶ GH3 cells with or without H19 stable overexpression were injected subcutaneously into nude mice. The normal GH3 cell mice were divided into two groups that received either gavage with saline solution or cabergoline. b The xenograft tumour size was monitored every other day (volume = width² × length × 1/2). At the end of the experiment, the xenograft tumours were dissected, photographed and weighed (c). d 4E-BP1 phosphorylation is more intensely suppressed in xenograft tumours treated with H19 than those treated with CAB. Some of the tumour tissues were homogenized in lysis buffer with a tissue homogenizer; then, the samples were centrifuged, and the supernatant was collected and used for immunoblot analysis with antibodies against total 4E-BP1 and phosphorylated 4E-BP1 Thr37/36 and Thr70. Tubulin was used as the loading control. e H19 suppresses GH3 cell proliferation more potently than CAB treatment (50 μM) in vitro. f, g CAB induces H19 expression in GH3 cells in both a dose-dependent (f) and time-dependent (g) manner. Total RNA was extracted from GH3 cells, followed by CAB treatment at different CAB doses and periods of time. qRT-PCR was employed to measure the H19 expression level in GH3 cells after CAB treatment. Relative H19 expression was normalized to actin expression. Error bars are the mean ± SEM values, ***p < 0.001