BRCA1 deficiency in ovarian cancer is associated with alteration in expression of several key regulators of cell motility – A proteomics study

Functional loss of expression of breast cancer susceptibility gene 1(BRCA1) has been implicated in genomic instability and cancer progression. There is emerging evidence that BRCA1 gene product (BRCA1) also plays a role in cancer cell migration. We performed a quantitative proteomics study of EOC patient tumor tissues and identified changes in expression of several key regulators of actin cytoskeleton/cell adhesion and cell migration (CAPN1, 14-3-3, CAPG, PFN1, SPTBN1, CFN1) associated with loss of BRCA1 function. Gene expression analyses demonstrate that several of these proteomic hits are differentially expressed between early and advanced stage EOC thus suggesting clinical relevance of these proteins to disease progression. By immunohistochemistry of ovarian tumors with BRCA1+/+ and BRCA1null status, we further verified our proteomic-based finding of elevated PFN1 expression associated with BRCA1 deficiency. Finally, we established a causal link between PFN1 and BRCA1-induced changes in cell migration thus uncovering a novel mechanistic basis for BRCA1-dependent regulation of ovarian cancer cell migration. Overall, findings of this study open up multiple avenues by which BRCA1 can potentially regulate migration and metastatic phenotype of EOC cells.

[1]  George M Yousef,et al.  Profilin-1 expression is associated with high grade and stage and decreased disease-free survival in renal cell carcinoma. , 2015, Human pathology.

[2]  J. Haugh,et al.  Profilin-1 serves as a gatekeeper for actin assembly by Arp2/3-dependent and -independent pathways. , 2015, Developmental cell.

[3]  Xuejuan Gao,et al.  Direct interaction of 14-3-3ζ with ezrin promotes cell migration by regulating the formation of membrane ruffle. , 2014, Journal of molecular biology.

[4]  M. B. Major,et al.  Modulation of Kaposi's Sarcoma-Associated Herpesvirus Interleukin-6 Function by Hypoxia-Upregulated Protein 1 , 2014, Journal of Virology.

[5]  F. Lin,et al.  14-3-3τ Promotes Breast Cancer Invasion and Metastasis by Inhibiting RhoGDIα , 2014, Molecular and Cellular Biology.

[6]  J. Condeelis,et al.  Profilin-1 downregulation has contrasting effects on early vs late steps of breast cancer metastasis , 2014, Oncogene.

[7]  T. Fehm,et al.  Macrophage Capping Protein CapG Is a Putative Oncogene Involved in Migration and Invasiveness in Ovarian Carcinoma , 2014, BioMed research international.

[8]  D. Perrotti,et al.  Protein phosphatase 2A: a target for anticancer therapy. , 2013, The Lancet. Oncology.

[9]  J. Fei,et al.  Expression of cofilin 1 is positively correlated with the differentiation of human epithelial ovarian cancer. , 2012, Oncology letters.

[10]  Scott D. Hansen,et al.  Differential remodeling of actin cytoskeleton architecture by profilin isoforms leads to distinct effects on cell migration and invasion. , 2012, Cancer cell.

[11]  Yong Ho Bae,et al.  Molecular insights on context-specific role of profilin-1 in cell migration , 2012, Cell adhesion & migration.

[12]  I. Ellis,et al.  Calpain system protein expression in basal-like and triple-negative invasive breast cancer , 2012, Annals of oncology : official journal of the European Society for Medical Oncology.

[13]  E. Friauf,et al.  Profilin1 is required for glial cell adhesion and radial migration of cerebellar granule neurons , 2012, EMBO reports.

[14]  T. Conrads,et al.  Standardization of a sample preparation and analytical workflow for proteomics of archival endometrial cancer tissue. , 2011, Journal of proteome research.

[15]  Hung-Ming Wang,et al.  Molecular Chaperones as a Common Set of Proteins That Regulate the Invasion Phenotype of Head and Neck Cancer , 2011, Clinical Cancer Research.

[16]  E. Coene,et al.  A novel role for BRCA1 in regulating breast cancer cell spreading and motility , 2011, The Journal of cell biology.

[17]  Sonja Gillen,et al.  Reduced expression of the membrane skeleton protein beta1-spectrin (SPTBN1) is associated with worsened prognosis in pancreatic cancer. , 2010, Histology and histopathology.

[18]  Yong Ho Bae,et al.  Profilin1 regulates PI(3,4)P2 and lamellipodin accumulation at the leading edge thus influencing motility of MDA-MB-231 cells , 2010, Proceedings of the National Academy of Sciences.

[19]  J. Lachuer,et al.  Gene expression profiles of human melanoma cells with different invasive potential reveal TSPAN8 as a novel mediator of invasion , 2010, British Journal of Cancer.

[20]  M. van de Rijn,et al.  Gross genomic alterations and gene expression profiles of high- grade serous carcinoma of the ovary with and without BRCA1 inactivation , 2010, BMC Cancer.

[21]  B. Karlan,et al.  Gene expression profile of BRCAness that correlates with responsiveness to chemotherapy and with outcome in patients with epithelial ovarian cancer. , 2010, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[22]  G. Mills,et al.  Somatic mutations in BRCA1 and BRCA2 could expand the number of patients that benefit from poly (ADP ribose) polymerase inhibitors in ovarian cancer. , 2010, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[23]  R. Bohle,et al.  Immunhistochemical analysis for expression of calpain 1, calpain 2 and calpastatin in endometrial cancer. , 2010, Anticancer research.

[24]  Lin-Yu Lu,et al.  The role of BRCA1 in DNA damage response , 2010, Protein & Cell.

[25]  A. Wells,et al.  Both actin and polyproline interactions of profilin-1 are required for migration, invasion and capillary morphogenesis of vascular endothelial cells. , 2009, Experimental cell research.

[26]  Timothy Y. Huang,et al.  Reactive oxygen species regulate a slingshot-cofilin activation pathway. , 2009, Molecular biology of the cell.

[27]  O. Medalia,et al.  Profilin 1 is required for abscission during late cytokinesis of chondrocytes , 2009, The EMBO journal.

[28]  Hua Guo,et al.  14-3-3zeta overexpression defines high risk for breast cancer recurrence and promotes cancer cell survival. , 2009, Cancer research.

[29]  Feng Zhang,et al.  PALB2 Links BRCA1 and BRCA2 in the DNA-Damage Response , 2009, Current Biology.

[30]  W. Foulkes,et al.  BRCA1 mutations contribute to cell motility and invasion by affecting its main regulators , 2008, Cell cycle.

[31]  R. Eeles,et al.  "BRCAness" syndrome in ovarian cancer: a case-control study describing the clinical features and outcome of patients with epithelial ovarian cancer associated with BRCA1 and BRCA2 mutations. , 2008, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[32]  J. Langowski,et al.  Invasive breast cancer cells exhibit increased mobility of the actin‐binding protein CapG , 2007, International journal of cancer.

[33]  A. Stojadinovic,et al.  HYOU1/Orp150 expression in breast cancer. , 2007, Medical science monitor : international medical journal of experimental and clinical research.

[34]  Rochelle L. Garcia,et al.  Functional Characterization of a Novel BRCA1-Null Ovarian Cancer Cell Line in Response to Ionizing Radiation , 2007, Molecular Cancer Research.

[35]  Lijun Xu,et al.  Suppression of Cancer Cell Migration and Invasion by Protein Phosphatase 2A through Dephosphorylation of μ- and m-Calpains* , 2006, Journal of Biological Chemistry.

[36]  P. Roy,et al.  Silencing profilin-1 inhibits endothelial cell proliferation, migration and cord morphogenesis , 2006, Journal of Cell Science.

[37]  Eithne Costello,et al.  Pancreatic cancer cells overexpress gelsolin family-capping proteins, which contribute to their cell motility , 2006, Gut.

[38]  J. Krischer,et al.  BRCA1 and BRCA2 mutations account for a large proportion of ovarian carcinoma cases , 2005, Cancer.

[39]  Rajendra K. Sharma,et al.  Overexpression of m-calpain in human colorectal adenocarcinomas. , 2004, Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology.

[40]  Kum Kum Khanna,et al.  BRCA1-BARD1 Complexes Are Required for p53Ser-15 Phosphorylation and a G1/S Arrest following Ionizing Radiation-induced DNA Damage* , 2004, Journal of Biological Chemistry.

[41]  A. Yu,et al.  The association of 14-3-3gamma and actin plays a role in cell division and apoptosis in astrocytes. , 2002, Biochemical and biophysical research communications.

[42]  H. Ozçelik,et al.  Epigenetic factors controlling the BRCA1 and BRCA2 genes in sporadic ovarian cancer. , 2002, Cancer research.

[43]  J. Fox,et al.  Calpain Cleaves RhoA Generating a Dominant-negative Form That Inhibits Integrin-induced Actin Filament Assembly and Cell Spreading* , 2002, The Journal of Biological Chemistry.

[44]  Christos Sotiriou,et al.  Gene expression profiles of BRCA1-linked, BRCA2-linked, and sporadic ovarian cancers. , 2002, Journal of the National Cancer Institute.

[45]  P. Greer,et al.  Reduced Cell Migration and Disruption of the Actin Cytoskeleton in Calpain-deficient Embryonic Fibroblasts* , 2001, The Journal of Biological Chemistry.

[46]  Bo Xu,et al.  Involvement of Brca1 in S-Phase and G2-Phase Checkpoints after Ionizing Irradiation , 2001, Molecular and Cellular Biology.

[47]  B. Rosen,et al.  Prevalence and penetrance of germline BRCA1 and BRCA2 mutations in a population series of 649 women with ovarian cancer. , 2001, American journal of human genetics.

[48]  S. Wesselborg,et al.  The serine phosphatases PP1 and PP2A associate with and activate the actin‐binding protein cofilin in human T lymphocytes , 2000, European journal of immunology.

[49]  H Nojima,et al.  A truncated isoform of the PP2A B56 subunit promotes cell motility through paxillin phosphorylation , 2000, The EMBO journal.

[50]  C. Welter,et al.  Expression of calpain I messenger RNA in human renal cell carcinoma: Correlation with lymph node metastasis and histological type , 1999, International journal of cancer.

[51]  M. Nakao,et al.  The involvement of calpain-independent proteolysis of the tumor suppressor NF2 (merlin) in schwannomas and meningiomas , 1998, Nature Medicine.

[52]  Sean P. Palecek,et al.  Regulation of Cell Migration by the Calcium-dependent Protease Calpain* , 1997, The Journal of Biological Chemistry.

[53]  Ralph Scully,et al.  Dynamic Changes of BRCA1 Subnuclear Location and Phosphorylation State Are Initiated by DNA Damage , 1997, Cell.

[54]  H. Kajiyama,et al.  Spectrin αII and βII tetramers contribute to platinum anticancer drug resistance in ovarian serous adenocarcinoma , 2012, International journal of cancer.

[55]  R. Bohle,et al.  Immunhistochemical analysis for expression of calpain 1, calpain 2 and calpastatin in ovarian cancer. , 2011, European journal of gynaecological oncology.

[56]  Alan Wells,et al.  Calpain proteases in cell adhesion and motility. , 2005, International review of cytology.

[57]  木村 麗新.  The involvement of calpain-dependent proteolysis of the tumor suppressor NF2 (merlin) in schwannomas and meningiomas , 1999 .