HE4 suppresses the expression of osteopontin in mononuclear cells and compromises their cytotoxicity against ovarian cancer cells

Abstract

Ovarian cancers are known to evade immunosurveillance and to orchestrate a suppressive immune microenvironment. Here we examine the role of human epididymis protein 4 (HE4), an ovarian cancer biomarker, in immune evasion. Through modified subtractive hybridization analyses we have characterized the gene targets of HE4 in human peripheral blood mononuclear cells (PBMCs), and established a preliminary mechanism for HE4-mediated immune failure in ovarian tumours. Upon exposure of purified PMBCs to HE4, osteopontin (OPN) and dual-specificity phosphatase 6 (DUSP6) emerged as the most suppressed and up-regulated genes, respectively. SKOV3 and OVCAR8, human ovarian carcinoma cell lines, exhibited enhanced proliferation in conditioned media from HE4-exposed PBMCs, an effect that was attenuated by the addition of recombinant OPN or OPN-inducible cytokines [interleukin (IL)-12 and interferon (IFN)-Ɣ]. Additionally, upon co-culture with PBMCs, HE4-silenced SKOV3 cells were found to be more susceptible to cytotoxic cell death. The relationship between HE4 and OPN was reinforced further through the analysis of serous ovarian cancer patient samples. In these biopsy specimens, the number of OPN T cells correlated positively with progression free survival (PFS) and inversely with serum HE4 level. Taken together, these findings show that HE4 enhances ovarian cancer tumorigenesis by compromising OPN-mediated T cell activation.

Inhibition of DUSP6 sensitizes ovarian cancer cells to chemotherapeutic agents via regulation of ERK signaling response genes

“…Immunohistochemical staining of an ovarian cancer microarray (US Biomax, OV802a) was performed as previously described [50], using antibodies for HE4 (Santa Cruz, sc-293473) and DUSP6 (MyBioSource, MBS8516662). Confocal microscopy was performed by an independent imaging technician at the Rhode Island Hospital Digital Imaging Core Facility with a Nikon C1si confocal (Nikon Inc. Mellville, NY, USA).…”

Gene Expression Profile of Active HE4 Stimulation in Epithelial Ovarian Cancer Cells: Microarray Study and Comprehensive Bioinformatics Analysis

“…In recent years, a growing number of investigations have gradually found that HE4 promotes cell proliferation, adhesion, invasion, migration, and chemoresistance in ovarian cancer [20][21][22][23][24][25][26][42][43][44][45][46][47]. It was found that HE4 overexpression or rHE4 treatment in EOC cells resulted in upregulation of many transcripts coding for extracellular matrix proteins, including LAMC2, LAMB3, SERPINB2 and GREM1; moreover, in cells overexpressing HE4 or exposed to rHE4 in culture medium, the protein levels of LAMC2 and LAMB3 were continously increased, and in the presence of bronectin, the focal adhesions were elevated in cells treated with rHE4 [22].…”

“…It was known that ovarian cancer participates in evading immunosurveillance and orchestrating a suppressive immune microenvironment, a series of studies by James NE, et al [23,44,45] found that, upon exposure of puri ed human peripheral blood mononuclear cells(PBMCs) to HE4, osteopontin (OPN) and DUSP6 appeared as the most inhibited and upregulated genes; the proliferation of human ovarian carcinoma cells in conditioned media from HE4-exposed PBMCs was enhanced, while the effect was attenuated by adding recombinant OPN or OPN-inducible cytokines (IL-12 and IFN-γ); HE4 can compromise both OPN-mediated T cell activation [44] and cytotoxic CD8 + /CD56 + cells through upregulation of self-produced DUSP6 [45], thus promoting the tumorigenesis of ovarian cancer [23,44,45,48]. Other researchers found that HE4 promotes carcinogenesis of ovarian cancer by combining with histone deacetylase 3 (HDAC3) to activate PI3K/AKT pathway [46], and that HE4 knockdown suppresses the invasive cell growth and malignant progress of ovarian cancer by inhibiting JAK/STAT3 pathway [24].…”

Gene expression profiles following active HE4 stimulation in epithelial ovarian cancer cells: microarray study and comprehensive bioinformatics analysis

“…Ovarian cancer participates in the evasion of immunosurveillance and orchestrates a suppressive immune microenvironment. A series of studies by James et al [23,44,45] showed that upon exposure of puri ed human peripheral blood mononuclear cells to HE4, osteopontin (OPN) and DUSP6 were the most downregulated and upregulated genes, respectively. The proliferation of human ovarian carcinoma cells in conditioned media from HE4exposed peripheral blood mononuclear cells was enhanced, whereas this effect was attenuated by adding recombinant OPN or OPN-inducible cytokines (interleukin-12 and interferon-γ).…”

“…The proliferation of human ovarian carcinoma cells in conditioned media from HE4exposed peripheral blood mononuclear cells was enhanced, whereas this effect was attenuated by adding recombinant OPN or OPN-inducible cytokines (interleukin-12 and interferon-γ). HE4 can compromise both OPN-mediated T cell activation [44] and the activity of cytotoxic CD8 + /CD56 + cells by upregulating self-produced DUSP6 [45], thus promoting the tumorigenesis of ovarian cancer [23,44,45,48]. Other researchers revealed that HE4…”