Oncogenic Protein Kinase D3 Regulating Networks in Invasive Breast Cancer

Abstract

Protein Kinase D3 (PRKD3) functions as an important oncogenic driver in invasive breast cancer, which is the leading cause of women mortality. However, PRKD3 regulating network is largely unknown. In this study, we systematically explored PRKD3 regulating networks via investigating phosphoproteome, interactome and transcriptome to uncover the molecular mechanism of PRKD3 in invasive breast cancer. Using iTRAQ, 270 proteins were identified as PRKD3 regulated phosphoproteins from 4619 phosphosites matching 3666 phosphopeptides from 2016 phosphoproteins with p-value <0.005. Transcriptome analysis using affymetrix microarray identified 45 PRKD3 regulated genes, in which 20 genes were upregulated and 25 genes were downregulated with p-value <0.005 upon silencing PRKD3. Using Co-IP in combination of MS identification, 606 proteins were identified to be PRKD3 interacting proteins from 2659 peptides. Further network analysis of PRKD3 regulated phosphoproteins, interacting proteins and regulated genes, reveals 19 hub nodes, including ELAVL1, UBC and BRCA1. UBC was recognized as the most common hub node in PRKD3 regulating networks. The enriched pathway analysis reveals that PRKD3 regulates pathways contributing to multiple cancer related events, including cell cycle, migration and others. Enrichment of cell cycle and cell mobility related pathways across PRKD3 networks, explained the observations that depletion of oncogenic PRKD3 led to alternation of cell cycle and decrease of cell migration ability. Taken together, our current study provided valuable information on the roles as well as the molecular mechanisms of PRKD3 in invasive breast cancer.

Protein Kinase D3 promotes the cell proliferation by activating the ERK1/c‐MYC axis in breast cancer

“…2,12 In order to study PRKD3 signalling pathway in breast cancer, CRISPR/Cas9 technology was applied for knocking out PRKD3 in the cells. 2,12 In order to study PRKD3 signalling pathway in breast cancer, CRISPR/Cas9 technology was applied for knocking out PRKD3 in the cells.…”

“…[2][3][4][5][6][7] Additionally, PRKD3 has been suggested to be up-regulated and involved in mediating the survival, differentiation, migration and proliferation of the triple-negative breast cancer cells. [2][3][4][5][6][7] Additionally, PRKD3 has been suggested to be up-regulated and involved in mediating the survival, differentiation, migration and proliferation of the triple-negative breast cancer cells.…”

“…[8][9][10] Diacylglycerol (DAG) has been found to bind to the cysteine-rich domain of PRKD3 and played important roles in PRKD3 activation via PKC signalling. 2,12 However, the interaction mechanisms among PRKD3, ERK1 and c-MYC in cancer cells has not been well-investigated since only one prostate cancer study suggested PRKD3-activated ERK1/2 is involved in tumour growth. The integrated phosphoproteomic and transcriptomic analysis in the previous studies showed that PRKD3 may play important roles in a variety of the cancer-related pathways.…”

“…Protein Kinase D3 has been implicated in a variety of functions in broad ranges of cancer types. [2][3][4][5][6][7] Additionally, PRKD3 has been suggested to be up-regulated and involved in mediating the survival, differentiation, migration and proliferation of the triple-negative breast cancer cells. [8][9][10] Diacylglycerol (DAG) has been found to bind to the cysteine-rich domain of PRKD3 and played important roles in PRKD3 activation via PKC signalling.…”

“…ERK1/ c-MYC axis was identified as a major pivot in PRKD3-regulated pathways in tumour cells. 2,12 However, the interaction mechanisms among PRKD3, ERK1 and c-MYC in cancer cells has not been well-investigated since only one prostate cancer study suggested PRKD3-activated ERK1/2 is involved in tumour growth. 13 The extracellular signal-regulated kinase 1/2 (ERK1/2) can be activated and/ or up-regulated by upstream kinases, transcription factors and/ or differentiation factor.…”