ABSTRACT
Prostate cancer (PCa) is one of the major public health problems in Western countries. Recently, the TMPRSS2:ERG gene fusion, which results in the aberrant expression of the transcription factor ERG, has been shown to be the most common gene rearrangement in PCa. Previous studies have determined the contributions of this fusion in PCa disease initiation and/or progression in vitro and in vivo. In this study on TMPRSS2:ERG regulation in PCa, we used an androgen receptor and TMPRSS2:ERG fusion double-negative PCa cell model: PC3c. In three cell clones with different TMPRSS2:ERG expression levels, ectopic expression of the fusion resulted in significant induction of cell migration and invasion in a dose-dependent manner. In agreement with this phenotype, high-throughput microarray analysis revealed that a set of genes, functionally associated with cell motility and invasiveness, were deregulated in a dose-dependent manner in TMPRSS2:ERG-expressing cells. Importantly, we identified increased MMP9 (Metalloproteinase 9) and PLXNA2 (Plexin A2) expression in TMPRSS2:ERG-positive PCa samples, and their expression levels were significantly correlated with ERG expression in a PCa cohort. In line with these findings, there was evidence that TMPRSS2:ERG directly and positively regulates MMP9 and PLXNA2 expression in PC3c cells. Moreover, PLXNA2 upregulation contributed to TMPRSS2:ERG-mediated enhancements of PC3c cell migration and invasion. Furthermore, and importantly, PLXNA2 expression was upregulated in metastatic PCa tumors compared with localized primary PCa tumors. This study provides novel insights into the role of the TMPRSS2:ERG fusion in PCa metastasis.
Subject(s)
Matrix Metalloproteinase 9/metabolism , Nerve Tissue Proteins/metabolism , Oncogene Proteins, Fusion/metabolism , Prostatic Neoplasms/metabolism , Receptors, Cell Surface/metabolism , Cell Line, Tumor , Cell Movement , Cell Proliferation , Gene Expression Regulation , Humans , Lymphatic Metastasis , Male , Oncogene Proteins, Fusion/genetics , Phenotype , Prostatic Neoplasms/pathology , TranscriptomeABSTRACT
Two ELISA assays were developed to test the reactivity of soluble sHLAs with anti-HLA class I mAbs of IgG or IgM isotype. A panel of 40 different alleles of sHLA antigens was produced using 57 lymphoblastoid B-cell lines, which had been generated from class-I-phenotyped PBLs. Using 14 mAbs, the expression of 13 different sHLA antigens (sHLA-A2, -A3, -A11, -A24, -A29, -B7, -B8, -B13, -B14, -B27, -B44, -B57, and -B58) by 43 different cell lines was confirmed. In addition, the expected absence of these alleles from the culture supernatant of 11 cell lines was confirmed. Cross-reactivities of mAbs observed by microlymphocytotoxicity assays were also detected by ELISA. The results of this extensive analysis confirmed previous results demonstrating that sHLA class I typing by ELISA correlated with HLA class I cell typing by microlymphocytotoxicity [1-3]. Furthermore, additional information about the fine specificities of two mAbs was obtained. An anti-B27/44 IgM mAb appeared to react only with sHLA-B44 but not with sHLA-B27; mAb CR11-351, previously reported to react with HLA-A2, 28, bound also to sHLA-A1, -3, -11, and -A24.
