Silencing of the cytokine receptor TNFRSF13B: A new therapeutic target for triple-negative breast cancer

Cytokine. 2020 Jan:125:154790. doi: 10.1016/j.cyto.2019.154790. Epub 2019 Aug 7.

Abstract

Background: TNFRSF13B, TACI, is a member of the TNF receptor superfamily; it plays a key role in cancer cell proliferation and progression.

Method: Influence of silencing of human cytokine receptors on cell viability was screened by Luminescent Cell Viability Assay, after transfection of the siRNA library to find the maximum cell death superhits in both triple-negative MDA-MB-231 and double-positive MCF7 breast cells. The mode of cell death was investigated by dual DNA fluorescence staining. The expression of mRNAs of TACI, BAFF, BAFF-R, and APRIL was explored by qPCR. Immunocytofluorescence analysis was used to evaluate changes in TACI, Bcl-2, TNFR2, cyclin-D2, and PCNA. NF-kB p65, cell cycle, and necrosis/apoptosis (late and early) were analyzed by flow cytometry.

Results: TACI is the most potent cytotoxic superhit resulted from high-throughput screening of the siRNA library, in both types of cells. Our findings indicated that silencing receptor TACI in both types of breast cancer cells led to significant cell death, after different intervals from siRNA transfection. Cell death mediators (TNFR2, Bcl-2, and NF-κB) were significantly decreased after TACI silencing. The key factors for cell division (Cyclin-D2 and PCNA) were significantly increased in silenced cells of both types but the cell cycle was arrested before the completion of mitosis. Expression of BAFF, BAFF-R and APRIL mRNA in TACI-silenced cells showed significant upregulation in MDA-MB-231 cells, while only BAFF-R and APRIL showed significant downregulation in MCF7 cells.

Conclusion: TACI silencing can be a new and promising therapeutic target for mesenchymal-stem like triple-negative breast cancer subtype.

Keywords: APRIL; BAFF; BAFF-R; Bcl-2; NF-κB; TACI; TNBC; TNFR2; TNFRSF13B; siRNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / genetics*
  • B-Cell Activating Factor / genetics
  • B-Cell Activating Factor / metabolism
  • B-Cell Activation Factor Receptor / genetics
  • B-Cell Activation Factor Receptor / metabolism
  • Cell Cycle / genetics*
  • Cell Survival / genetics
  • Cyclin D2 / genetics
  • Cyclin D2 / metabolism
  • Down-Regulation
  • Female
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Gene Expression Regulation, Neoplastic / genetics*
  • Gene Silencing
  • High-Throughput Screening Assays
  • Humans
  • MCF-7 Cells
  • NF-kappa B / genetics
  • NF-kappa B / metabolism
  • Necrosis / metabolism
  • Proliferating Cell Nuclear Antigen / genetics
  • Proliferating Cell Nuclear Antigen / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • RNA, Small Interfering
  • Real-Time Polymerase Chain Reaction
  • Transmembrane Activator and CAML Interactor Protein / genetics
  • Transmembrane Activator and CAML Interactor Protein / metabolism*
  • Triple Negative Breast Neoplasms / genetics
  • Triple Negative Breast Neoplasms / metabolism*
  • Tumor Necrosis Factor Ligand Superfamily Member 13 / genetics
  • Tumor Necrosis Factor Ligand Superfamily Member 13 / metabolism
  • Up-Regulation

Substances

  • B-Cell Activating Factor
  • B-Cell Activation Factor Receptor
  • BCL2 protein, human
  • CCND2 protein, human
  • Cyclin D2
  • NF-kappa B
  • Proliferating Cell Nuclear Antigen
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Small Interfering
  • TNFRSF13B protein, human
  • TNFRSF13C protein, human
  • TNFSF13B protein, human
  • Transmembrane Activator and CAML Interactor Protein
  • Tumor Necrosis Factor Ligand Superfamily Member 13