Patient-based cross-platform comparison of oligonucleotide microarray expression profiles

Schlingemann, Joerg; Habtemichael, Negusse; Ittrich, Carina; Toedt, Grischa; Hj, Kramer; Hambek, Markus; Knecht, Rainald; Lichter, Peter; Stauber, Roland H.; Hahn, Meinhard

doi:10.1038/labinvest.3700293

Cited by 52 publications

(40 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Increased MRCKα expression has been reported in a number of cancer types, including adenocarcinoma (39), cutaneous squamous cell carcinoma (SCC) (40), in oral, hypopharyngeal, head and neck SCC, oral cavity and tongue carcinoma (41)(42)(43)(44)(45)(46)(47). In addition, increased MRCKα expression has also been identified in in vitro studies using U937 histiocytic lymphoma, MDA-MB 231 breast cancer, A549 lung cancer and PLB-985 myelocytic leukemia cell lines (46,47).…”

Section: Discussionmentioning

confidence: 92%

Afzelin exhibits anti-cancer activity against androgen-sensitive LNCaP and androgen-independent PC-3 prostate cancer cells through the inhibition of LIM domain kinase 1

et al. 2015

View full text Add to dashboard Cite

Abstract. Prostate cancer presents high occurrence worldwide. Medicinal plants are a major source of novel and potentially therapeutic molecules; therefore, the aim of the present study was to investigate the possible anti-prostate cancer activity of afzelin, a flavonol glycoside that was previously isolated from Nymphaea odorata. The effect of afzelin on the proliferation of androgen-sensitive LNCaP and androgen-independent PC-3 cells was evaluated by performing a water soluble tetrazolium salt-1 assay. In addition, the effect of afzelin on the cell cycle of the LNCaP and PC-3 prostate cancer cell lines was evaluated. Western blot analysis was performed to evaluate the effect of afzelin on the kinases responsible for the regulation of actin organization. Afzelin was identified to inhibit the proliferation of LNCaP and PC3 cells, and block the cell cycle in the G 0 phase. The anticancer activity of afzelin in these cells was determined to be due to inhibition of LIM domain kinase 1 expression. Thus, the in vitro efficacy of afzelin against prostate cancer is promising; however, additional studies on different animal models are required to substantiate its anticancer potential. IntroductionProstate cancer is the second most frequently diagnosed type of cancer and the sixth most common cause of cancer-associated mortality in males, worldwide (1). The typical treatment strategies of chemotherapy and radiotherapy have not provided significant survival benefits for patients with advanced prostate cancer, and the majority of available strategies are only palliative (2). Therefore, there is requirement for the prompt identification of novel molecules to treat the increasing number of prostate cancer cases, particularly cases that are resistant to current chemotherapeutic agents (3,4).Afzelin is a flavonol glycoside found in Nymphaea odorata, which has been identified to inhibit the growth of breast cancer cells by stimulating apoptosis (5). In addition, afzelin has been demonstrated to scavenge superoxide anion radicals in RAW264.7 cells (6). The structure of this compound is shown in Fig. 1.In eukaryotic cells, the actin cytoskeleton is essential for mediating various biological functions, including providing the structural framework of the cell, and driving cellular motility and division. In particular, dynamic reorganization of the actomyosin cytoskeleton and remodelling of the extracellular matrix drive the multistep process of tumor cell metastasis. Tumor cells are able to cross tissue boundaries into the blood and lymphatic systems and migrate to distal regions of the body. Therefore, cancer therapy has recently focused on gaining a comprehensive understanding of the biological processes that regulate actin organization (7).Rho GTPase family proteins are key regulators of the actin cytoskeleton and, with the aid of various target proteins, maintain the tight regulation of normal cell growth and differentiation (8-11). Eukaryotic cells exhibit a predisposition to rapid and uncontrollable growth following genomic a...

show abstract

Section: Discussionmentioning

confidence: 92%

Afzelin exhibits anti-cancer activity against androgen-sensitive LNCaP and androgen-independent PC-3 prostate cancer cells through the inhibition of LIM domain kinase 1

et al. 2015

View full text Add to dashboard Cite

show abstract

“…Plasmids were verified by sequence analysis as described previously (21). Oligonucleotides used for PCR amplification and cloning are listed in supplemental Table S1.…”

Section: Methodsmentioning

confidence: 99%

“…mRNA preparation and cDNA synthesis from tumor tissue was performed as described previously (21). Cloning of the Taspase1 coding sequence into expression vectors pc3, pc3-GFP, pc3-BFP, and pc3-mCherry using BamHI/EcoRI or BamHI/NheI restriction sites, respectively, allowed the expression of Taspase1, alone or as a fusion with fluorescent proteins as described previously (17,22).…”

Section: Methodsmentioning

confidence: 99%

Cell-based Analysis of Structure-Function Activity of Threonine Aspartase 1

Bier

Knauer

Klapthor

et al. 2011

Journal of Biological Chemistry

105

View full text Add to dashboard Cite

Taspase1 is a threonine protease responsible for cleaving intracellular substrates. As such, (de)regulated Taspase1 function is expected not only to be vital for ordered development but may also be relevant for disease. However, the full repertoires of Taspase1 targets as well as the exact biochemical requirements for its efficient and substrate-specific cleavage are not yet resolved. Also, no cellular assays for this protease are currently available, hampering the exploitation of the (patho)-biological relevance of Taspase1. Here, we developed highly efficient cell-based translocation biosensor assays to probe Taspase1 trans-cleavage in vivo. These modular sensors harbor variations of Taspase1 cleavage sites and localize to the cytoplasm. Expression of Taspase1 but not of inactive Taspase1 mutants or of unrelated proteases triggers proteolytic cleavage and nuclear accumulation of the biosensors. Employing our assay combined with scanning mutagenesis, we identified the sequence and spatial requirements for efficient Taspase1 processing in liquid and solid tumor cell lines. Collectively, our results defined an improved Taspase1 consensus recognition sequence,, allowing the first genome-wide bioinformatic identification of the human Taspase1 degradome. Among the 27 most likely Taspase1 targets are cytoplasmic but also nuclear proteins, such as the upstream stimulatory factor 2 (USF2) or the nuclear RNA export factors 2/5 (NXF2/5). Cleavage site recognition and proteolytic processing of selected targets were verified in the context of the biosensor and for the full-length proteins. We provide novel mechanistic insights into the function and bona fide targets of Taspase1 allowing for a focused investigation of the (patho)biological relevance of this type 2 asparaginase.By cleaving proteins, proteases are involved in the control of a large number of key physiological processes such as development, metabolism, tissue remodeling, cell proliferation, and apoptosis (1-3). Protease signaling therefore differs from the majority of other signaling pathways by being mostly irreversible (3). Protease signaling is strictly regulated, and the deregulation of protease activity can contribute to various pathologies, including cancer (3).The human Taspase1 gene encodes a protein of 420 amino acids (aa), 4 which is the proenzyme of Taspase1. It belongs to a family of enzymes possessing an asparaginase-2 homology domain. In the MEROPS database, Taspase1 is found as T02.004, classifying this protein as a class PB, subclass PB(T), and T2 family protease. In contrast to the other cis-active type 2 asparaginases, such as amidohydrolases, L-asparaginase, and glycosylasparaginase, only Taspase1 is able to cleave other substrates in trans (4). Therefore, Taspase1 represents a distinct class of proteolytic enzymes. Taspase1-mediated cleavage of proteins follows distinct aspartate residues, suggesting that Taspase1 evolved from hydrolyzing asparagines and glycosylasparagines to recognize a conserved peptide motif with an aspartate at the P1 ...

show abstract

“…Hybridization of 15 mg labeled cRNA was done on HG-U133A Affymetrix microarrays (Affymetrix, Santa Clara, CA) as described previously. 36 All arrays were globally scaled to a target value of 1000, and intracellular PLK1 mRNA values (Affymetrix Average Difference Units) were derived using Microarray Suite 5.0 software.…”

Section: Microarray Expression Analysismentioning

confidence: 99%

Polo-Like Kinase 1 as Predictive Marker and Therapeutic Target for Radiotherapy in Rectal Cancer

Rödel

Keppner

Capalbo

et al. 2010

The American Journal of Pathology

View full text Add to dashboard Cite

The ability to predict tumor sensitivity toward radiotherapy may significantly impact the selection of patients for preoperative combined-modality therapy. The aim of the present study was to test the predictive value of Polo-like kinase 1 (PLK1) in rectal cancer patients and to investigate whether PLK1 plays a direct role in mediating radiation sensitivity. PLK1 expression was evaluated by immunohistochemistry (n ‫؍‬ 76) or Affymetrix HG133 microarray (n ‫؍‬ 20) on pretreatment biopsies of patients with advanced rectal cancer. Expression was correlated with both tumor regression in the resected specimen and long-term clinical outcome. Furthermore, we used small interfering RNAs (siRNAs) to down-regulate PLK1 expression in colorectal cancer cells and analyzed the effects of PLK1-specific siRNAs by Western blot and quantitative real-time PCR analysis, FACScan analysis, caspase 3/7 assays, and colony-forming assays. We observed that increased PLK1 protein expression was significantly related to a poorer tumor regression and a higher risk of local recurrence in uniand multivariate analysis. A significant decrease of PLK1 expression by siRNAs in combination with ionizing radiation induced an increased percentage of apoptotic cells and increased caspase 3/7 activity. Furthermore, enhanced G 2 -M levels, decreased cellular viability, and reduced clonogenic survival were demonstrated, indicating a radiosensitizing effect of PLK1 depletion. Therefore, PLK1 may be a novel predictive marker for radiation response as well as a promising therapeutic target in rectal cancer patients. Recent data from preoperative radiochemotherapy (RCT) in rectal cancer indicate a broad variety in tumor responses, ranging from pathological complete response with no viable tumor cells left to virtually no tumor regression at all.1 Despite uniform treatment protocols, this observed heterogeneity in treatment response is most probably caused by the individual genetic accouterment of the tumor. Current research has focused on molecular markers involved in the resistance toward anticancer therapy including key elements of the apoptotic and mitotic pathways.2,3 Among these factors, the serine/threonine kinase Polo-like kinase 1 (PLK1) has emerged as prognostic factor and as promising target for cancer therapy, 4,5 because it is a key regulator for the mitotic progression of mammalian cells, 6 -8 and it is overexpressed in all human cancers analyzed up to date. -12Several preclinical approaches including the use of PLK1 antibodies, dominant-negative forms of PLK1, antisense oligonucleotides, small interfering RNA (siRNA) and small molecule inhibitors have consistently revealed cell cycle arrest at G 2 -M, and inhibition of tumor proliferation both in vitro and in vivo associated with the induction of apoptosis. [13][14][15][16][17][18][19] A growing number of small molecule inhibitors is currently under clinical investigation. 20 -24 PLK1 kinase activity is inhibited by DNA damaging agents, in an ataxia telangiectasia mutated or Rad3-related kin...

show abstract

Patient-based cross-platform comparison of oligonucleotide microarray expression profiles

Cited by 52 publications

References 40 publications

Afzelin exhibits anti-cancer activity against androgen-sensitive LNCaP and androgen-independent PC-3 prostate cancer cells through the inhibition of LIM domain kinase 1

Afzelin exhibits anti-cancer activity against androgen-sensitive LNCaP and androgen-independent PC-3 prostate cancer cells through the inhibition of LIM domain kinase 1

Cell-based Analysis of Structure-Function Activity of Threonine Aspartase 1

Polo-Like Kinase 1 as Predictive Marker and Therapeutic Target for Radiotherapy in Rectal Cancer

Contact Info

Product

Resources

About