Abstract

Abstract The accumulation of numerous chromosomal aberrations is a hallmark characteristic of invasive breast cancer (IBC). It is thus critical to find specific chromosomal aberrations that may drive breast cancer initiation and progression, particularly those that are drivers of the genomic instability. By integrative analysis of genomic data from The Cancer Genome Atlas (TCGA), we have nominated Tousled-Like Kinase 2 (TLK2) as a candidate oncogene target for IBC. Ultra-high resolution copy-number data from TCGA suggest that TLK2 was amplified in approximately 10% of IBC, which are more frequent in luminal B than luminal A tumors. These data also revealed that TLK2-high breast tumors harbor significantly more global copy-number aberrations than the TLK2 low-tumors. TLK2 is a nuclear serine/threonine kinase that serves as a cell cycle checkpoint regulating chromatin assembly, and in response to DNA damage, TLK2 can be inactivated by phosphorylation through the ATM-dependent pathway. We thus postulated that up-regulation of TLK2 may help cancer cells to surpass the cell cycle checkpoint upon DNA damage, which may promote genomic instability in breast cancer cells. To analyze the effect of TLK2 overexpression, we introduced an inducible expression vector containing the TLK2 open-reading frame into the MCF10A benign epithelial cells and the T47D breast cancer cells. As shown by fluorescence activated cell sorting (FACS), ectopic expression of TLK2 in these cells overcame the cell cycle checkpoint in response to the DNA damage induced by gamma irradiation. gamma-H2AX foci formation revealed increased DNA double strand breaks and prolonged DNA repair window after irradiation in the TLK2 overexpressing cells. Interestingly, immunofluorescence localized TLK2 to the nucleus of breast cancer cells, which was markedly enriched in DNA double-strand break foci (or gamma-H2AX foci). To examine the effect of TLK2 knockdown, the MCF7 and MDAMB361 breast cancer cells overexpressing TLK2 were transfected with different TLK2 siRNAs. TLK2 inhibition led to G1/S cell cycle arrest (FACS analysis), profound decrease in cell growth (MTT assay), and enhanced apoptosis (Annexin V assay) in the MCF7 and MDAMB361 breast cancer cells over-expressing TLK2. Together, this study supports the role of TLK2 deregulation in the increased genomic instability of TLK2-high breast cancers, and suggests its potential as an attractive therapeutic target. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P3-04-04.

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