Overexpression of NNT-AS1 Activates TGF-β Signaling to Decrease Tumor CD4 Lymphocyte Infiltration in Hepatocellular Carcinoma

Figure 3

Six immune subtypes of cancer, C1 (wound healing), C2 (IFN-γ dominant), C3 (inflammation), C4 (lymphocyte depletion), C5 (immune silencing), and C6 (TGF-β dominance). There were no samples with the C5 subtype in the LIHC data. (a) Six immune subtypes of cancer, C1 (wound healing), C2 (IFN-γ dominant), C3 (inflammation), C4 (lymphocyte depletion), C5 (immune silencing), and C6 (TGF-β dominance). There were no samples with the C5 subtype in the LIHC data. The levels of NNT-AS1 in different HCC immune subtypes. (b) Pearson correlation analysis of the levels of NNT-AS1 and genes involved in TGF-β signaling and other genes involved in interferon signaling and the immune reaction. (c) RT-qPCR determination of the NNT-AS1 levels in three cell lines (HL-7702, HepG2, and Huh7, repeat three times). (d) NNT-AS1 expression was confirmed by RT-qPCR after transfection of siRNA targeting TGF-β, TGFBR1, or SMAD5 into HepG2 cells (repeat three times). (e, f) Human recombinant TGF-β activated the TGF-β signaling pathway and increased the expression of NNT-AS1. SB431542 inhibited the TGF-β signaling pathway and decreased the expression of NNT-AS1 (repeat 6 times). (g) The correlations of NNT-AS1 with SMAD5 and TGFBR1 in 15 HCC patients were analyzed by RT-qPCR.

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