https://orcid.org/0000-0003-4111-7603
Figures & data
Figure 1 The YBX1 expression profiles in different type of human cancers.
Notes: (A) Increased or decreased mRNA levels of YBX1 in datasets of various cancers compared with normal tissues via Oncomine database. (B) Representative datasets of BRCA with decreased YBX1 expression compared with normal tissues in Oncomine. (C) The expression levels of YBX1 in different tumor types analyzed by TIMER database. (D) The protein levels of YBX1in different type of cancer cell lines. **P<0.01, ***P< 0.001 compared with corresponding normal tissues.
Abbreviations: BRCA, breast cancer; RPPA, reverse phase protein arrays; TIMER, the Tumor Immune Estimation Resource database; YBX1, Y-box binding protein 1.
Figure 2 Subgroup analysis of YBX1 mRNA level in BRCA by UALCAN database.
Notes: Boxplot for relative mRNA levels of YBX1 in normal tissues and BRCA samples with different (A) genders; (B) age stages; (C) histologic subtypes; (D) cancer stages; (E) TP53 mutation status; (F) molecular subtypes; (G) triple-negative BRCA subtypes; (H) nodal metastasis status. *P< 0.05, **P<0.01, ***P< 0.001 compared with normal tissues.
Abbreviations: BRCA, breast cancer; IDC, invasive ductal carcinoma; ILC, invasive lobular carcinoma; INOS, infiltrating– not otherwise specified; TCGA, The Cancer Genome Atlas; TP53, tumor protein 53; YBX1, Y-box binding protein 1.
Figure 3 The correlation between YBX1 expression and clinicopathological parameters in BRCA.
Notes: (A) The clustering of clinicopathological parameters according to the YBX1 mRNA level using cBioPeortal database. (B) Immunohistochemical staining of YB1 in BRCA samples with different molecular subtypes. (C) The different protein expression of YB1 in human breast epithelial cells and aseries of BRCA cell lines. *P<0.05 and ***P<0.001 vs control group.
Abbreviations: BRCA, breast cancer; cBioPortal, the cBio Cancer Genomics Portal database; ns, not significant; TNBC, triple-negative breast cancer; YBX1, Y-box binding protein 1.
Table 1 Correlation of YBX1 Expression and Clinical Prognosis in Breast Cancer with Different Clinicopathological Parameters by Kaplan–Meier Plotter
Figure 4 Survival curves comparing the high and low expression of YBX1 in BRCA by Kaplan–Meier plotter database.
Notes: (A) Survival curves of OS in the entire BRCA cohort and different subtypes; (B) Survival curves of RFS in the entire BRCA cohort and different subtypes.
Abbreviations: BRCA, breast cancer; HR, hazard ratio; OS, overall survival; RFS, relapse-free survival; YBX1, Y-box binding protein 1.
Table 2 Correlation Analysis Between YBX1 and Markers of Immune Infiltrates via TIMER
Figure 5 Correlation of YBX1 expression with immune cell infiltration level in (A) the entire BRCA cohort; (B) the luminal A subtype; (C) the luminal B subtype; (D) the HER2-enriched subtype; and (E) the triple-negative subtype by TIMER database.
Abbreviations: BRCA, breast cancer; TIMER, the Tumor Immune Estimation Resource database; YBX1, Y-box binding protein 1.
Figure 6 Verification of the correlation between YBX1 and immune cell infiltration in BRCA.
Notes: (A) Infiltration levels of different type of macrophages in samples from GSE1456 dataset with different YBX1 expression level. (B) Kaplan–Meier curves from the TIMER database showing the impacts of YBX1 expression and M1/M2 infiltration on survival of BRCA patients. (C) Ascatter plot of correlation between YBX1 and various genes especially those associated with immunosuppression (annotated in red) in BRCA. (D) Correlation of YBX1 with IDO1, PDCD1, CTLA4 and LAG3 in BRCA verified by cBioPortal database. (E) Immunohistochemical staining of IDO1 and CTLA4 in luminal BRCA samples with different YB1 expression levels. **P<0.01 and ***P<0.001 vs corresponding YB1 high-expressing groups.
Abbreviations: BRCA, breast cancer; cBioPortal, the cBio Cancer Genomics Portal database; CTLA4, cytotoxic T-lymphocyte associated protein 4; IDO1, indoleamine 2,3-dioxygenase 1; LAG3, lymphocyte activating 3; NES, normalized enrichment score; HR, hazard ratio; PDCD1 programmed cell death 1; TIMER, Tumor Immune Estimation Resource; TCGA, The Cancer Genome Atlas; YBX1, Y-box binding protein 1.
Figure 7 Effect of YBX1 knockdown on the cytotoxicity of macrophages and CD8+ T cells in co-culture system of BRCA cells and immunocytes.
Notes: (A) YBX1 was effectively knocked down by siRNAs in MCF-7, BT-474, and MDA-MB-231 cells. (B) BRCA cells with or without knockdown of YBX1 were cultured alone or co-cultured with peritoneal macrophages, and cell viability was evaluated. For luminal BRCA cells, namely MCF-7 and BT-474, the decrease of cell viability after YBX1 knockdown were more dramatic in co-cultures compared with the single culture system. However, no difference was observed for MDA-MB-231 TNBC cells. (C) IL-2 production by activated CD8+ Tcells co-cultured with BRCA cells transfected with YBX1 siRNA or scrambled negative control siRNA. YBX1 knockdown significantly increased IL-2 secretion in co-cultures of CD8+ Tcells with MCF-7 or BT-474 cells but not in the co-culture with MDA-MB-231 cells. (D) IFN-γ production by activated CD8+ Tcells co-cultured with BRCA cells transfected with YBX1 or scrambled negative control siRNA. YBX1 knockdown significantly increased IFN-γ secretion in the co-cultures of CD8+ Tcells and MCF-7 or BT-474 cells but not in the co-culture with MDA-MB-231 cells. (E) The relative survival of BRCA cells represented by the release of LDH in co-cultures of CD8+ Tcells and BRCA cells with or without YBX1 knockdown. YBX1 depletion markedly reduced the survival of MCF-7 and BT-474 cells but not MDA-MB-231 cells. **P<0.01, ***P<0.001 compared with corresponding negative control groups.
Abbreviations: BRCA, breast cancer; IFN-γ, interferon-γ; IL-2, interleukin-2; LDH, lactate dehydrogenase; NC, negative control; YBX1, Y-box binding protein 1.
