图1 泛癌种中SLC1A5的表达情况

Fig1 Differential expressions of SLC1A5 across different cancer types. A: SLC1A5 expression in normal tissues. B: SLC1A5 expression in tumor cell lines. C: Expression status of SLC1A5 gene in paired tumor and non-tumor tissues across 22 cancer types from TCGA. D: Analysis of SLC1A5 expression in 33 cancer types with their corresponding normal tissues in GTEx dataset as controls. *P<0.05, **P<0.01, *** P<0.001.

图2 SLC1A5在多种癌症中高表达

Fig.2 SLC1A5 is highly expressed in various cancers. A: Expression and distribution of SLC1A5 in different cancer cell lines (Scale bar=10 μm); B-F: Comparison of SLC1A5 gene expression between normal and tumor tissues from TCGA databases (left), SLC1A5 protein expression between normal and tumor tissues from CPTAC (middle), and immunohistochemistry images of normal and tumor tissues from HPA (right, Scale bar=100 μm). SLC1A5 expression at both the mRNA and protein levels was significantly higher in colorectal cancer (B), lung cancer (C), and endometrial cancer (D). There was no significant difference in SLC1A5 expression between tumor and non-tumor in breast cancer (E). SLC1A5 gene expression was slightly upregulated in renal cancer, but was significantly downregulated in the tumor tissues at the protein level (F). *P<0.05, ***P<0.001.

图3 SLC1A5在各种癌症类型中高表达

Fig.3 SLC1A5 is highly expressed in various cancer types. Comparison of SLC1A5 gene expression between normal and tumor tissues from TCGA databases (left), SLC1A5 protein expression between normal and tumor tissues from CPTAC (middle), and immunohistochemistry images of normal and tumor tissues from HPA (right, Scale bar=100 μm). ***P<0.001.

图4 SLC1A5在不同癌种不同病理分期及淋巴转移与否的肿瘤组织中的表达水平

Fig.4 Expression levels of SLC1A5 gene in cancers in realtion to pathological stages and status of lymphatic metastasis. A: Expression levels of SLC1A5 mRNA in different pathological stages of HNSC, KIRC, KIRP, LIHC, PAAD, TGCT, and THCA. B: Expression levels of SLC1A5 mRNA analyzed in relation to lymphatic metastasis in HNSC, KIRC, LIHC, and THCA. *P<0.05, **P<0.01, ***P<0.001.

图5 SLC1A5表达与不同肿瘤生存预后的关系

Fig.5 Correlation between SLC1A5 expression and survival outcomes of different cancers. A: GEPIA2 tool was used for analysis of overall survival (A) and disease-survival (B) for 33 cancers in TCGA in relation to SLC1A5 expression. The survival map and survival curves with statistical significance are shown.

图6 SLC1A5表达水平与肿瘤微环境及肿瘤免疫细胞浸润水平的相关性

Fig.6 Correlation of SLC1A5 expression levels with tumor microenvironment and levels of tumor immune cell infiltration. A: Three tumors with the highest correlation coefficients between SLC1A5 expression and the tumor microenvironment. B: Correlation coefficients between SLC1A5 expression and tumor infiltration level of different immune cells in 33 cancer types. *P<0.05; **P<0.01, ***P<0.001, ****P<0.0001.

图7 SLC1A5表达与免疫相关基因的共表达

Fig.7 Co-expression of SLC1A5 expression and immune-related genes. Correlations of SLC1A5 expression with immunosuppressive genes (A), immune activation genes (B), chemokines receptor (C), and chemokines (D) are presented.

图8 SLC1A5表达与不同亚型巨噬细胞的相关性

Fig.8 Correlation between SLC1A5 expression and different subtypes of macrophage. A: Relationship between SLC1A5 expression and different subtypes of macrophage. B: Three tumors with the highest correlation coefficients between SLC1A5 expression and M2 macrophages. C: Survival map illustrating the correlation between different subtypes of macrophage and overall survival in different cancers. D: Kaplan-Meier survival analysis of LGG (left) and LIHC (right) patients with different SLC1A5 expression levels stratified by levels of infiltrating M2 macrophages.

图9 过表达SLC1A5促进肝癌增殖

Fig.9 Overexpression of SLC1A5 promotes proliferation of HCC cells. A: Expression of SLC1A5 in HCC cell lines detected by Western blotting. LE: Long exposure; SE: Short exposure. B: Verification of SLC1A5 knockout and overexpression efficiency in different HCC cell lines using Western blotting. C: Effect of RNA interference of SLC1A5 on HCC cell proliferation assessed using CCK-8 assay. D: Effect of SLC1A5 overexpression on HCC cell proliferation assessed using by CCK-8 assay. E: Overexpression efficiency of SLC1A5 in HCC-LM3 cells examined by Western blotting (upper). Examination of subcutaneous tumors in nude mice (bottom). F, G: Proliferation curve of subcutaneous tumor (F) and tumor weight (G) in nude mice. H, I: Expressions of SLC1A5 and Ki-67 in subcutaneous tumors detected by immunohistochemistry. *P<0.05, **P<0.01, ***P<0.001.

图10 SLC1A5促进M2型巨噬细胞极化并抑制T细胞功能

Fig.10 SLC1A5 promotes M2 polarization of macrophages and induces CD8+ T cell dysfunction. A: Correlation analysis between SLC1A5 expression and molecular markers of M2 macrophages. B: M0 cells co-cultured with HCC cells transfected with SLC1A5-specific siRNA or negative control siRNA and induced to polarize into M2 phenotype using IL-4 (20 ng/mL). Interference efficiency of SLC1A5 and expressions of M2 markers (IL-10, TGFB1, CSF1R and CXCL-1) detected using qRT-PCR. C: M0 cells co-cultured with SLC1A5-overpressing HCC-LM3 cells and induced to polarize into M2 phenotype using IL-4 (20 ng/mL). The overpression efficiency of SLC1A5 and the expression of M2 markers (IL-10, TGFB1, CSF1R and CXCL-1) were detected using qRT-PCR. D, E: M0 cells co-cultured with HCC cells with SLC1A5 knockdown (D) and overexpression (E) and induced into the M2 phenotype using IL-4 (20 ng/mL). The culture medium was used to treat activated Jurkat T cells for 48 h, and IFN-γ secretion was detected by ELISA. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.