The activation of EP300 by F11R leads to EMT and acts as a prognostic factor in triple-negative breast cancers

Abstract

Cancer progression is influenced by junctional adhesion molecule (JAM) family members. The relationship between JAM family members and different types of cancer was examined using The Cancer Genome Atlas dataset. mRNA levels of the F11R (F11 receptor) in tumours were inversely correlated to the expression of JAM-2 and JAM-3. This relationship was unique to breast cancer (BCa) and was associated with poor prognosis (p = 0.024, hazard ratio = 1.44 [1.05-1.99]). A 50-gene molecular signature (prediction analysis of microarray 50) was used to subtype BCa. F11R mRNA expression significantly increased in human epidermal growth factor receptor 2 (HER2)-enriched (p = 0.0035) and basal-like BCa tumours (p = 0.0005). We evaluated F11R protein levels in two different compositions of BCa subtype patient tissue array cohorts to determine the relationship between BCa subtype and prognosis. Immunohistochemistry staining revealed that a high F11R protein level was associated with poor overall survival (p < 0.001; Taipei Medical University [TMU] cohort, p < 0.001; Kaohsiung Veterans General Hospital [KVGH] cohort) or disease-free survival (p < 0.001 [TMU cohort], p = 0.034 [KVGH cohort]) in patients with BCa. Comparison of F11R levels in different subtypes revealed the association of poor prognosis with high levels of F11R among luminal (p < 0.001 [TMU cohort], p = 0.027 [KVGH cohort]), HER2 positive (p = 0.018 [TMU cohort], p = 0.037 [KVGH cohort]), and triple-negative (p = 0.013 [TMU cohort], p = 0.037 [KVGH cohort]) BCa. F11R-based RNA microarray analysis and Ingenuity Pathway Analysis were successful in profiling the detailed gene ontology of triple-negative BCa cells regulated by F11R. The EP300 transcription factor was highly correlated with F11R in BCa (R = 0.51, p < 0.001). By analysing these F11R-affected molecules with the L1000CDs datasets, we were able to predict some repurposing drugs for potential application in F11R-positive BCa treatment.

Keywords: EP300; F11R; breast cancer; survival curve; transcriptomics.

Figure 1

Distribution of JAM members within BCa. (A) Expression of F11R in cancer types. Analysed data from TIMER 2.0 website. (B) Heatmap illustrating the expression of JAM members in BCa. The data were obtained from XENA website. (C) Boxplots illustrating the expression of the JAM gene family in the BCa normal and tumour groups. Data were analysed using the GEPIA 2.0 website (*p < 0.05; **p < 0.01; ***p < 0.001).

Figure 2

F11R and JAM‐2/JAM‐3 are inversely related to BCa prognosis. (A) Kaplan–Meier survival curves for JAM members in BCa with recurrence‐free survival. Data were analysed using the Kaplan–Meier plotter website. (B) HRs for JAM members with different BCa prognosis features under different clinical scenarios. The data were obtained from the PrognoScan website. (C) Distribution of JAM members in the different types of BCa. The data were analysed using the XENA website (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0001).

Figure 3

Relationship between F11R protein levels and prognosis in BCa subtype classification. (A) Grading of immunohistochemical staining for F11R between tumour and non‐tumour. (B) Correlation between expression of F11R and OS or DFS in different BCa tissues. (C) OS according to F11R and different BCa subtype classifications. *HR was not determined since all the cases in the low F11R group are censored.

Figure 4

Prediction of molecular biological events using F11R‐based transcriptomics in TNBC cells. (A) Venn diagram of significant changes in targets following microarray analysis of the F11R overexpression model in CAL120 and HDQ‐P1 cells. (B) Networks identified by IPA of F11R‐overexpressing cell models of BCa cells. (C) Potential signalling pathways predicted using IPA that may be affected by F11R overexpression. Data were analysed using IPA software.

Figure 5

The F11R/EP300 axis is involved in the regulation of cell motility in TNBC. (A) Correlation of selected EMT‐related molecules with F11R in BCa. (B) EMT‐selected molecules and F11R gene as a signature of BCa. (C) Upstream effectors of EP300 after the overexpression of F11R. (D) Relationship between EP300 and downstream effectors in BCa. Data were analysed using IPA software.

Figure 6

Simulation of drug repurposing based on F11R genetic profiles. (A) Illustration of the correlation between gene expression and drug candidates in a heatmap. (B) Tyrphostin AG 1478 is a potential candidate to inhibit TGFB1. Data were analysed using the L1000CDs website.