Increased expression of SRp40 affecting CD44 splicing is associated with the clinical outcome of lymph node metastasis in human breast cancer

Background: During the malignant transformation of breast tissues, the pre-mRNA precursor splicing of specific genes can be flexibly regulated, leading to the formation of different forms or amounts of mRNA in response to the cellular microenvironment, and is frequently associated with cell tumorigenesis and may even cause tumor metastasis. Regulation of pre-mRNA splicing by serine-arginine (SR)-rich phosphoprotein is hypothesized to be associated with tumor cell metastasis. Methods: We enrolled 55 breast cancer patients (32 with lymph node metastasis; LNM) with paired tissue samples consisting of cancerous and tumor-adjacent normal portions and assayed these tissues for gene expression of the SR family using quantitative real-time RT-PCR (qRT-PCR), then evaluated an association with LNM of breast cancer. Furthermore, we examined whether increased expression of a specific SR gene was associated with the presence of specific CD44 spliced variants using qualitative reverse-transcription PCR (RT-PCR). Results: Support for our hypothesis came from the observations that breast tumor tissues displayed higher level of SRp40 expression as compared with the paired non-cancerous tissues, which manifested the significant association between increased SRp40 expression and LNM (OR=4.48, 95% CI, 1.08-19.50, P=0.018). In addition, the primary tumors of breast with increased expression of SRp40 gene were associated with the presence of the large CD44 inclusion variants, CD44v2, CD44v3, CD44v5, and CD44v6 (P<0.05). Conclusion: Increased expression of SRp40 can be detected in breast tumor tissues with a high degree of sensitivity, and that higher expression of SRp40 closely correlates with alternative pre-mRNA splicing of CD44, which may serve as an earlier marker in predicting the risk to breast cancer patients of developing LNM. (c) 2007 Elsevier B.V. All rights reserved.