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Using 2D and 3D in vitro experiments as well as in vivo models, we show that USP5 is important for pancreatic cancer growth. X in Western blot analyses in response to doxycycline-induced USP5 knockdown. A p value of less than 0. In parallel, clone 3 was used to establish xenograft tumors in vivo by subcutaneously implanting cells mixed with matrigel in athymic nude mice. These polyubiquitinated proteins may then hinder cell growth via different mechanisms in different model systems.

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For details, please see Suppl. Ubiquitin and proteasomes in transcription. USP as a predictive and prognostic factor following neoadjuvant therapy in braiesh with breast cancer.

CA Cancer J Clin.

Samples of normal pancreatic tissue were obtained from healthy donors. Correlation between spontaneous metastatic potential and brsjesh I collagenolytic activity in a human pancreatic cancer cell line SUIT-2 and sublines.

Journal List Oncotarget v. Chemoresistance in pancreatic cancer is driven by stroma-derived insulin-like growth factors. Our results are consistent with the hypothesis that loss of USP5 leads to accumulation of DNA damage, which in turn leads to accumulation of p27, cell cycle pertubation, and increased apoptosis rates.

Mouse xenograft assay 2×10 6 cells in 0.

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Table 1 List of putative druggable target genes. USP5 knockdown in pancreatic cancer cells leads to accumulation of polyubiquitinated proteins Ubwhereas monomeric ubiquitin Ub is diminished. X, as well as increased levels of p21 and p27 and decreased levels of Cyclin D1 in the absence of USP5 function. Informed consent in writing was obtained from all patients prior to using tissue samples. The majority of genes were targeted by two or more individual shRNA expression constructs average: Similar to the results from transient RNAi experiments, we also observed upregulation of p27 and increased levels of phos.

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For data evaluation, the software FlowJo ver. Since p53 is either mutationally inactivated or deleted in the vast majority of PDAC cases as well as all cell lines used in our experimentsour data adds another layer to the functionality of this protein.

Regulation of gene expression by the ubiquitin-proteasome system. Possibly this difference reflects a combined effect of the unspecific cellular stress of transfection and the parallel loss of USP5 function in the transient siRNA experiments. Daviet L, Colland F. Deubiquitinase inhibition as a cancer therapeutic strategy. Among brajeeh candidate genes identified in this screen was the deubiquitinase USP5, barjesh subsequent gene expression analyses demonstrated to be significantly upregulated in primary human pancreatic cancer tissues.

Swatek KN, Komander D.

Other Cyclin proteins were unaffected. Cell cycle analyses of PaTuT cells with USP5 knockdown revealed significantly elevated proportions of cells in the G 1 -phase of the cell cycle compared to control siRNA treated or untreated cells, while the proportion of cells in S-phase 3f reduced accordingly. Unless stated otherwise, transient transfection was always done with 1×10 5 cells per well in a 6-well plate format.

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Contributed by Author contributions B. We did not observe any systematic differences in the expression levels of USP5 among cancer cell lines bfajesh different origin brjaesh between the cancer cells and the non-transformed cell-line HEK Supplementary Figure 1A. Cells were collected at t 0 and t endbarcode sequences were amplified from genomic DNA and subjected to high-throughput sequencing and compared for their abundance. Hoheisel2 Thomas M.

Pancreatic cancer is the fourth leading cause of cancer-related deaths in the United States [ 29 ] and is projected to become the second leading cause by [ 30 ]. This article has been cited by brajesj articles in PMC. In fact, USP5 itself has been reported to have a tumor inhibiting role in different disease models.

Whole-exome sequencing of pancreatic cancer defines genetic diversity and therapeutic targets. Comput Methods Programs Biomed. There is an increasing body of evidence suggesting that Brsjesh represent a new class of promising drug targets that can be targeted more specifically than the proteasome due to their distinct association with particular genetic or biochemical pathways, especially in diseases like cancer [ 1623 ].