Fig. 1

HMGA1 enhanced oncogenic progression of pCCA cells. a Left: Heatmap of HMG gene expression profiles in eight pairs of pCCA tissues and adjacent normal tissues. Right: FPKM of HMGA1 in pCCA and adjacent normal tissues were shown. Data were were analyzed by paired t tests (b) HMGA1 mRNA expression in 18 paired pCCA and adjacent normal duct tissues, detected by qRT-PCR. P value was calculated with paired t tests. c Up: HMGA1 expression in four randomly-selected pairs of pCCA tissues (T) and adjacent normal tissues (N), detected by Western blot. Bottom: quantification of bands in the left panel. d Representative images of immunohistochemical staining for low/high expression of HMGA1 in the tissue microarray (top: 100× magnification, scale bar, 200 μm; bottom: 400× magnification, scale bar, 50 μm). e Correlation between HMGA1 expression and clinicopathological features (Chi-square tests). f Kaplan-Meier survival analysis in patients with pCCA with low (n = 35) and high (n = 71) HMGA1 expression (cut-off: 77.4). g HMGA1 expression in human cholangiocarcinoma cell lines, gallbladder carcinoma cell lines, and intrahepatic bile duct cell line HiBEpiC. (H) HMGA1 was silenced in QBC939 and overexpressed in FRH0201, and the effects of HMGA1 on pCCA cell proliferation were measured by CCK-8 assays. i effects of HMGA1 on colony formation in QBC939 and FRH0201 cells after knockdown in QBC939 or overexpression in FRH0201. j QBC939 cells with stable HMGA1 knockdown (up) or overexpression (bottom) were subcutaneously injected to nude mice for xenograft model (n = 6/group). k Tumor volumes in xenografts established using cells infected with lentivirus carrying scrambled shRNA, shHMGA1, empty vector, or GV492-HMGA1. l,m The effects of HMGA1 on migration and invasion were detected with wound-healing assay(l) and transwell assay(m) after silencing HMGA1 in QBC939 or overexpressing HMGA1 in FRH0201 cells. n Effects of HMGA1 on stemness was detected by 7-day-sphere formation activity with stable HMGA1-silencing or HMGA1-overexpressing QBC939 cells. o,p Effects of HMGA1 on EMT were detected by Western blot after silencing HMGA1 in QBC939(o) and overexpressing HMGA1 in FRH0201 cells(p). *P < 0.05, **P < 0.01, and ***P < 0.001 compared with the corresponding control group. Data are shown as means ±S.D., and statistical significance was analyzed with one-way ANOVA(h and k) or T-test. Analyzed data were from three independent experiments, and each subgroup was performed at least in triplicate (c,g, h,i, l-p)