SETD3 acts as a prognostic marker in breast cancer patients and modulates the viability and invasion of breast cancer cells

Abstract

In several carcinomas, the SET Domain Containing 3, Actin Histidine Methyltransferase (SETD3)is associated with oncogenesis. However, there is little knowledge about the role of SETD3 in the progression and prognosis of breast cancer. In this study, we first analyzed the prognostic value of SETD3 in breast cancer patients using the database of the public Kaplan-Meier plotter. Moreover, in vitro assays were performed to assess the role of SETD3 in the viability and capacity of invasion of human breast cancer cell lines. We observed that the high expression of SETD3 was associated with better relapse-free survival (RFS) of the whole collective of 3,951 patients, of Estrogen Receptorpositive, and of Luminal A-type breast cancer patients. However, in patients lacking expression of estrogen-, progesterone-and HER2-receptor, and those affected by a p53-mutation, SETD3 was associated with poor RFS. In vitro analysis showed that SETD3 siRNA depletion affects the viability of triple-negative cells as well as the cytoskeletal function and capacity of invasion of highly invasive MDA-MB-231 cells. Interestingly, SETD3 regulates the expression of other genes associated with cancer such as β-actin, FOXM1, FBXW7, Fascin, eNOS, and MMP-2. Our study suggests that SETD3 expression can act as a subtype-specific biomarker for breast cancer progression and prognosis.MTT proliferation assay. SETD3 depleted cells or controls (5 × 10 3 ) were plated in 96-well plates with DMEM medium (without phenol red) (Gibco ® , cat. No. 31053028, Germany) with FCS and incubated for 96 hours. After the incubation time, the medium was removed completely and cells were incubated with 20 µl/well of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) (cat. No. M2128-1G), at 5 mg/ml for 4 hours, at 37 °C. Subsequently, 100 µl of Stopping buffer, pH 4.7 composed of 10% (w/v) sodium dodecyl sulfate (SDS) (cat. No. 3599286) and 50% (v/v) N,) was added to stop the reaction and dissolve the formazan crystals. The absorbance was measured in a VersaMax ® Microplate Reader (Molecular Devices, Sunnyvale, CA, USA) at a wavelength of 595 nm. The proliferation of the control cells was defined as 100%. Results were derived from three independent sets of triplicate experiments. MTT, SDS, and N,N-Dimethyl formamide were from Sigma-

Epigenetic Modifier SETD8 as a Therapeutic Target for High-Grade Serous Ovarian Cancer

“…Other SETD-domain protein methyltransferases, such as SETD3, have also been considered as therapeutic targets for cancer. For example, it has been reported that SETD3 negatively correlates with prognosis in breast cancers [ 29 ]. However, further clarification is required with regard to the role of this protein in HGSOC.…”

Prognostic significance of hedgehog signaling network‐related gene expression in breast cancer patients

“…The publically available gene expression database Kaplan‐Meier plotter (KM plotter) (https://kmplot.com) was used to analyze the prognostic value of the hedgehog signaling related‐genes in the relapse‐free survival of breast cancer patients 30 . The establishment of the database of the gene expression data and survival information was performed as were previously described in Espinoza‐Sánchez et al, 23 Hassan et al, 24 and Györffy et al 30 Patients were stratified by ER, progesterone receptor, HER2 expression, and lymph node status, as well as for the molecular classification, and grading. We analyzed the expression of some genes of the Hh pathway and visualized their correlation to survival by generating Kaplan‐Meier survival plots.…”

“…For example, we have recently shown that altered expression of the NF-κB pathway and the methyltransferase SETD3 are associated with subtype-specific poor survival of breast cancer patients. 23,24 However, the role of the Hh pathway in the initiation and progression of breast cancer is not well understood. GLI2 and PTCH1 play an important role in the development of the mammary gland in murine models.…”

“…Several signaling pathways and epigenetic factors contribute to the progression of breast cancer. For example, we have recently shown that altered expression of the NF‐κB pathway and the methyltransferase SETD3 are associated with subtype‐specific poor survival of breast cancer patients 23,24 . However, the role of the Hh pathway in the initiation and progression of breast cancer is not well understood.…”

Initial Identification of UDP-Glucose Dehydrogenase as a Prognostic Marker in Breast Cancer Patients, Which Facilitates Epirubicin Resistance and Regulates Hyaluronan Synthesis in MDA-MB-231 Cells

“…To assess the prognostic value of UGDH and other functionally linked genes, the online tool kmplot.com [35], which allows a meta-analysis of gene expression in relation to breast cancer patient survival, was employed [36]. Gene expression data was obtained through microarray analysis of widely used arrays of the GEO database and converted into Kaplan-Meier plots.…”