Fig. 2
From: HMGA1 augments palbociclib efficacy via PI3K/mTOR signaling in intrahepatic cholangiocarcinoma
HMGA1 promoted the transcription and expression of CCND1.
A, 39 genes were filtered out among the datasets of shHMGA, EMT and PluriNet signature genes.
B-C, 8 genes were screened from the top lists of the KEGG and the GO enrichment analysis and evaluated in RBE.
D, CCND1 mRNA levels in 18 pairs of iCCAs and adjacent normal bile duct tissues were detected with qRT-PCR and showed as log (CCND1 Tumor/CCND1Non-tumor).
E, Correlation between HMGA1 and CCND1 mRNA ratio in fresh iCCA tissues calculated by mRNA (tumor)/mRNA (adjacent tissue).
F-G, qRT-PCR and WB showed that HMGA1 expression regulated the expression of CCND1 in HUCCT1 and RBE cells.
H, HMGA1 promoted the transcription of CCND1 in both HUCCT1 and RBE cells and 293 T cells. The transcriptional activity of CCND1 was detected with luciferase assays.
I, The expression and localization of HMGA1 and CCND1 in HUCCT1 cells were detected with immunofluorescence. The co-localization of HMGA1 and CCND1 was attenuated after HMGA1 knockdown. Scale bar 20 μm.
J-K, WB showed that the PI3K signal pathway was regulated by HMGA1.
L-N, HMGA1-silenced or HMGA1-overexpressing HUCCT1 or RBE cells were incubated in PI3K agonist 740 Y-P (30 µM, 24 h) or PI3K inhibitor PF-04691502 (0.5 µM, 48 h), and CCND1 expression was detected with WB. In HMGA1-overexpressing HUCCT1 cells, CCND1 was knocked down, and OCT4, CD44 and (p)RB expression were detected with WB.
n.s. ,** and *** represented not signifificant, P < 0.01 and < 0.001, respectively. Analyzed data were from three independent experiments and shown as means ± SEM. Data were from three independent experiments and analyzed with the T-test(C, F-K). Spearman correlation analysis was performed in (E and I)