A functional role for the cancer disparity-linked genes, CRYβB2 and CRYβB2P1, in the promotion of breast cancer

Background In the USA, the breast cancer mortality rate is 41% higher for African-American women than non-Hispanic White women. While numerous gene expression studies have classified biological features that vary by race and may contribute to poorer outcomes, few studies have experimentally tested these associations. CRY beta B2 gene expression has drawn particular interest because of its association with overall survival and African-American ethnicity in multiple cancers. Several reports indicate that overexpression of the CRY beta B2 pseudogene, CRY beta B2P1, and not CRY beta B2 is linked with race and poor outcome. It remains unclear whether either or both genes are linked to breast cancer outcomes. This study investigates CRY beta B2 and CRY beta B2P1 expression in human breast cancers and breast cancer cell line models, with the goal of elucidating the mechanistic contribution of CRY beta B2 and CRY beta B2P1 to racial disparities. Methods Custom scripts for CRY beta B2 or CRY beta B2P1 were generated and used to identify reads that uniquely aligned to either gene. Gene expression according to race and tumor subtype were assessed using all available TCGA breast cancer RNA sequencing alignment samples (n = 1221). In addition, triple-negative breast cancer models engineered to have each gene overexpressed or knocked out were developed and evaluated by in vitro, biochemical, and in vivo assays to identify biological functions. Results We provide evidence that CRY beta B2P1 is expressed at higher levels in breast tumors compared to CRY beta B2, but only CRY beta B2P1 is significantly increased in African-American tumors relative to White American tumors. We show that independent of CRY beta B2, CRY beta B2P1 enhances tumorigenesis in vivo via promoting cell proliferation. Our data also reveal that CRY beta B2P1 may function as a non-coding RNA to regulate CRY beta B2 expression. A key observation is that the combined overexpression of both genes was found to suppress cell growth. CRY beta B2 overexpression in triple-negative breast cancers increases invasive cellular behaviors, tumor growth, IL6 production, immune cell chemoattraction, and the expression of metastasis-associated genes. These data underscore that both CRY beta B2 and CRY beta B2P1 promote tumor growth, but their mechanisms for tumor promotion are likely distinct. Conclusions Our findings provide novel data emphasizing the need to distinguish and study the biological effects of both CRY beta B2 and CRY beta B2P1 as both genes independently promote tumor progression. Our data demonstrate novel molecular mechanisms of two understudied, disparity-linked molecules.