Abstract
Autophagy is a catabolic process for degrading dysfunctional proteins and organelles, and closely associated with cancer cell survival under therapeutic, metabolic stress, hypoxia, starvation and lack of growth factors, contributing to resistance to therapies. However, the role of autophagy in breast cancer cells is not well understood. In the present study, we investigated the role of autophagy in highly aggressive and metastatic triple negative breast cancer (TNBC) and non-metastatic breast cancer cells and demonstrated that the knockdown of autophagy-related genes (LC3 and Beclin-1) inhibited autophagy and significantly suppressed cell proliferation, colony formation, migration/invasion and induced apoptosis in MDA-MB-231 and BT-549 TNBC cells. Knockdown of LC3 and Beclin-1 led to inhibition of multiple proto-oncogenic signaling pathways, including cyclin D1, uPAR/integrin-β1/Src, and PARP1. In conclusion, our study suggests that LC3 and Beclin-1 are required for cell proliferation, survival, migration and invasion, and may contribute to tumor growth and progression of highly aggressive and metastatic TNBC cells and therapeutic targeting of autophagy genes may be a potential therapeutic strategy for TNBC in breast cancer.
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This study was supported by The Scientific and Technological Research Council—TÜBİTAK—research Grant (Project number: 214S204).
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This study was funded by The Scientific and Technological Research Council—TÜBİTAK—research Grant (Grant number 214S204).
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Supplementary Figure 1. Expression of LC3 and Beclin-1 proteins in BT-549 TNBC cells
. Cells were lysed and expression of LC3-I, LC3-II and beclin1 proteins were determined by Western blot using specific antibodies β-actin was used as a loding control, Supplementary Figure 2. Effect of inhibition of autophagy by LC3 and Beclin-1 knockdown on cell viability of MCF7 cells. siRNA-mediated knockdown of autophagy gene did induced marked effect on cell proliferation/viability under normal (A) and starvation conditions (B). Proliferation/ cell viability was detected by an MTS assay, Supplementary Figure 3. Effect of inhibition of autophagy by LC3 and Beclin-1 knockdown on colony formation of MCF7 cells. siRNA-mediated knockdown of autophagy gene did not inhibit MCF7 cell proliferation and colony formation. MCF7 cells were transfected with indicated siRNAs and evaluated for colony formation by crystal violet staining. (PPTX 473 KB)
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Hamurcu, Z., Delibaşı, N., Geçene, S. et al. Targeting LC3 and Beclin-1 autophagy genes suppresses proliferation, survival, migration and invasion by inhibition of Cyclin-D1 and uPAR/Integrin β1/ Src signaling in triple negative breast cancer cells. J Cancer Res Clin Oncol 144, 415–430 (2018). https://doi.org/10.1007/s00432-017-2557-5
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DOI: https://doi.org/10.1007/s00432-017-2557-5