The fact that many biomarkers are used to categorize specific cancers has proven beneficial for people's understanding of cancer and for developing clinical diagnostic methods that allow for more effective treatment. Based on the use of markers such as ER, PR, and HER2, BRCA is currently classified as luminal, HER2-enriched, and TNBC. These subtypes, however, are not yet fully understood in terms of molecular regulation. Identifying specific markers to aid in diagnosis and treatment is therefore extremely important. In this study, we investigate the JAM family of tumors from a pan-cancer perspective through the use of open big data. Several types of cancer express the F11R gene in combination with the JAM family (Figure.1A), which has previously been reported as well [26, 27]. Notably, F11R was confirmed in this study to be elevated in BRCA expression as well as unique among other members of the JAM family in terms of its associated prognostic characteristics. In support of these findings, according to previous studies, F11R is related to many clinical prognoses associated with the BRCA gene [26, 27, 35]. Intriguingly, the results indicate that JAM-2 and JAM-3 act as inverse indicators of F11R. Comparing the pan-cancer analysis, BLCA, BRCA, CESC, LUSC, and UCEC, similar trends were seen (Figure.S1A). In particular, such a relationship is positively correlated with OS and a poor prognosis is associated with F11R gene deficiencies in BRCA patients (Figure.S1B). TCGA-BRCA data also indicated that in the normal case, F11R exhibited a lower distribution than JAM-2 or JAM-3 (Figure.1B). Comparing the differences between normal and tumor populations also revealed that in contrast to the results of JAM-2 or JAM-3, F11R was significantly higher in the tumor group than in the normal group. The clinical correlation between the JAM family and Recurrence Free Survival, in addition to OS, was similar to that seen in BRCA. A review of different clinical data also indicated a similar situation, suggesting that having the F11R results in a high chance of a RFS, disease specific survival or DFS of patients (Figure.2A,B), while JAM-2/JAM-3 is associated with low risk. However, it should be noted that the relationship between JAM family members and BRCA subtypes has not been systematically evaluated. In this study, a series of comparisons are made between different BRCA subtypes based on the expression level of the JAM genes. Both in-silico and IHC results showed that F11R was significantly increased in all subtypes, except in HER2-enriched and Basal-like subtypes (Figure.2C, 3B), we demonstrate that F11R is associated with OS with Luminal, HER2 and TNBC (Figure.3C). In contrast to F11R, patients with any BRCA subtype will have lower levels of JAM-2/JAM-3 expression. The same trends are also observed in patients with HER2-enriched or Basal-like tumors. However, according to the correlation, there is no correlation between the distributions of F11R and JAM-2/JAM-3 among the inverse indicators (Figure.S2). It is nonetheless noteworthy that there is a strong correlation between JAM-2/JAM-3 (R = 0.8). Despite this, no studies have demonstrated a possible relationship or biological function between JAM-2 or JAM-3.
Currently, TNBC is currently considered to be one of the most difficult subtypes to treat, and metastasis is currently considered to be one of the main causes Considering that there was no detailed analysis to profiling how F11R functions in TNBC cells, we therefore simulated the possible molecular mechanism between F11R and TNBC cells through microarray profiling analysis. Cells overexpressing F11R were shown to activate gene ontology related to cell growth and motility (Figure.4B). Analysis of Canonical pathways also revealed the major signaling mechanisms involved in F11R (Figure.4C). Based on the findings of this study, it is evident that F11R plays a role in several processes related to cell motility, including tight junction signaling, regulation of the EMT by growth factors, HER2 signaling in BRCA, and ER signaling, both of which are associated with BRCA tumorigenesis. It is consistent with recent studies suggesting that HER2-induced resistance to treatment is mediated by HER3 and is mediated by F11R in luminal and basal-like BRCA cell lines . Overexpression of F11R can also be observed in similar cells, regulating Integrin Signaling and influencing cell motility . Our observations are consistent with the results of this study (Figure.4C). Here, we demonstrate that F11R can exhibit consistent signaling regulation in both basal-like and mesenchymal-like TNBC cells. In light of these results, F11R appears to be a therapeutic target that may be cross-subtype applicable, in accordance with the IHC results.
In fact, F11R is involved in EMT-related signaling in cells, which is consistent with the features we have observed in pathological studies. On the molecular level, the EMT is regulated by growth factors via the F11R signaling pathway (Figure.5A), and molecules with high correlation with FSTL3 include GSK3B, MAPK1, SMAD4, PTPN11, OCLN, CDC42, MTQR, TGFBR1, RHOA, and IL6R (Figure.5B). Several of these molecules are indeed highly correlated with F11R in BRCA (Figure.5C) and have previously been implicated in BRCA progression [37–40]. It should be noted that these molecules may serve as cluster signatures and are highly correlated with F11R in BRCA. With a more detailed analysis of Upstream regulators, we identified a novel factor involved in F11R regulation, the EP300 (Figure.5D). EP300 expression is consistently linked to a wide range of biological processes, including cell growth, metastasis, and stemness properties of TNBC . An IPA analysis indicated that EP300 regulates RHOA, GSK3B, and TGFBR1. An analysis of correlations revealed a strong correlation between EP300 and F11R, RHOA, GSK3B and TGFBR1 (Figure.5E). Similar correlations were also observed among different subtypes of BRCA cell lines (Figure.S4). There have been a number of studies that have demonstrated the connection between RHOA, GSK3B, and TGFBR1 and the development of metastatic disease in BRCA [37–40]. This suggests that the crosstalk between F11R and TGFRB1 signaling may also be mediated by EP300. However, the precise mechanism needs to be determined through additional research.
As a result of our study, we suggest that the expression level of biomarker F11R is useful for the diagnosis and evaluation of the disease, especially in subtypes of BRCA. Moreover, as the interpretation between the molecular networks demonstrates, F11R participates in BRCA progression through molecular mechanisms, and can be used as a potential target for future drug design in BRCA therapy.