scholarly journals Specific and reliable detection of Myosin 1C isoform A by RTqPCR in prostate cancer cells

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5970 ◽  
Author(s):  
Aleena A. Saidova ◽  
Daria M. Potashnikova ◽  
Anna V. Tvorogova ◽  
Ivan V. Maly ◽  
Wilma A. Hofmann ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Mrna Expression ◽  
Cancer Cells ◽  
Cell Lines ◽  
Reference Genes ◽  
Critical Issue ◽  
Cell Detection ◽  
Mrna Levels ◽  
Prostate Cell ◽  
Prostate Cell Lines

Background Prostate cancer (PC) diagnostics and treatment often present a challenging task due to cancer subtype heterogeneity and differential disease progression in patient subgroups. Hence, the critical issue is finding a reliable and sensitive diagnostic and prognostic PC marker, especially for cases of biopsies with low percentages of cancer cells. Isoform A of myosin 1C was shown to be expressed in PC cells and responsible for their invasive properties, however, its feasibility for diagnostic purposes remains to be elucidated. Methods To verify the role of myosin 1C isoform A mRNA expression as a putative prostate cancer marker we performed RT qPCR normalized by three reference genes (GAPDH, YWHAZ, HPRT1) on PC3, RWPE-1, LNCaP and 22Rv1 cell lines. Myosin 1C isoform A detection specificity was confirmed by immunofluorescence staining, cancer and non-cancer prostate cell lines were immunophenotyped by flow cytometry. Results Median normalized mRNA expression level of myosin 1C isoform A in PC cells (PC3 and 22Rv1) is two orders of magnitude higher compared to RWPE-1 cells, which functionally correspond to benign prostate cells. Myosin 1C isoform A expression allows PC cell detection even at a dilution ratio of 1:1000 cancer to non-cancer cells. At the protein level, the mean fluorescence intensity of myosin 1C isoform A staining in PC3 nuclei was only twice as high as in RWPE-1, while the immunophenotypes of both cell lines were similar (CD44+/CD90-/CD133-/CD57-/CD24+-). Conclusions We report a distinct difference in myosin 1C isoform A mRNA levels in malignant (PC3) and benign (RWPE-1) prostate cell lines and suggest a combination of three reference genes for accurate data normalization. For the first time we provide an immunophenotype comparison of RWPE-1 and PC3 cells and demonstrate that RT qPCR analysis of MYO 1C A using appropriate reference genes is sufficient for PC detection even in low-abundance cancer specimens.

scholarly journals NCL Inhibition Exerts Antineoplastic Effects against Prostate Cancer Cells by Modulating Oncogenic MicroRNAs

Cancers ◽  
2020 ◽  
Vol 12 (7) ◽  
pp. 1861
Author(s):  
Tyler Sheetz ◽  
Joseph Mills ◽  
Anna Tessari ◽  
Megan Pawlikowski ◽  
Ashley E. Braddom ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cancer Cells ◽  
Cell Lines ◽  
Therapeutic Option ◽  
The United States ◽  
Mrna Levels ◽  
Advanced Stage ◽  
Castration Resistant Prostate Cancer ◽  
Single Chain

Prostate cancer (PCa) is the most frequently diagnosed cancer in men and second most common cause of cancer-related deaths in the United States. Androgen deprivation therapy (ADT) is only temporarily effective for advanced-stage PCa, as the disease inevitably progresses to castration-resistant prostate cancer (CRPC). The protein nucleolin (NCL) is overexpressed in several types of human tumors where it is also mislocalized to the cell surface. We previously reported the identification of a single-chain fragment variable (scFv) immuno-agent that is able to bind NCL on the surface of breast cancer cells and inhibit proliferation both in vitro and in vivo. In the present study, we evaluated whether NCL could be a valid therapeutic target for PCa, utilizing DU145, PC3 (CRPC), and LNCaP (androgen-sensitive) cell lines. First, we interrogated the publicly available databases and noted that higher NCL mRNA levels are associated with higher Gleason Scores as well as with recurrent and metastatic tumors. Then, using our anti-NCL scFv, we demonstrated that NCL is expressed on the surface of all three tested cell lines and that NCL inhibition results in reduced proliferation and migration. We also measured the inhibitory effect of NCL targeting on the biogenesis of oncogenic microRNAs such as miR-21, -221 and -222, which was cell context dependent. Taken together, our data provide evidence that NCL targeting inhibits the key hallmarks of malignancy in PCa cells and may provide a novel therapeutic option for patients with advanced-stage PCa.

scholarly journals CHARACTERIZATION OF COLLAGEN (IV) MRNA IN CELL LINES OF BREAST CANCER

2015 ◽  
Vol 77 (25) ◽  
Author(s):  
Afzan Mat Yusof ◽  
Mardhiah Mohammad ◽  
Sharifah Norbaizura Syed Bahrom ◽  
Syahirah Kaja Mohideen ◽  
Ridhwan Roshdi ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Mrna Expression ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Cancer Cell Lines ◽  
Collagen Iv ◽  
Breast Cancer Cell Lines ◽  
Mrna Levels

Breast cancer incidence rate has increased in the 5 recent years with 14% increases in mortality. The structural change in the collagen chain has led to alterations in the cancer cells. Various biological processes, such as differentiation or gene expression, are regulated through extracelullar matrix (ECM)[1]. The restructuring of the collagenous architecture in the hypoxic microenvironment may influence the invasive growth of the cancer cells. With the increased stress within the cell, the invasion of cancer cells into the ECM was triggered. This cell lines model would enable the exploration of the relationship between the extracellular matrices component and the tumor proliferation. The aim of this study is to characterize the collagen (IV) mRNA expression in the breast cancer cell.  Breast cancer (MCF7) cell lines were cultured and harvested upon confluent. The RNA was extracted from the cell lines and then the cDNA were synthesized. The collagen (IV) mRNA levels in breast cancer cell lines were measured using real time PCR and GAPDH was used as an internal control. The level of COL4A2 (IV) mRNA expression was higher compared with COL4A1 (IV) mRNA. The level of COL4A5 (IV) mRNA was reduced significantly in breast cancer cells lines. Overall, the expression of COL4A1-A6 (IV) was reduced. The reduced amount of collagen (IV) in breast cancer cell lines suggested that the collagen was restructured and this has triggered the tumor invasion into the ECM.

scholarly journals ZFP91: A Noncanonical NF-κB Signaling Pathway Regulator with Oncogenic Properties Is Overexpressed in Prostate Cancer

2016 ◽  
Vol 2016 ◽  
pp. 1-8 ◽  
Author(s):  
Lukasz Paschke ◽  
Karol Jopek ◽  
Marta Szyszka ◽  
Marianna Tyczewska ◽  
Agnieszka Ziolkowska ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cancer Cells ◽  
Signaling Pathway ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Biology ◽  
Prostate Cancer Cell ◽  
Cancer Cell Lines ◽  
Mrna Levels ◽  
Prostate Cancer Cell Lines

Novel molecular targets are being searched to aid in prostate cancer diagnosis and therapy. Recently, ZFP91 zinc finger protein has been found to be upregulated in prostate cancer cell lines. It is a potentially important oncogenic protein; however only limited data regarding its biological function and expression patterns are available. To date, ZFP91 has been shown to be a key factor in activation of noncanonical NF-κB signaling pathway as well as to be involved in HIF-1α signaling in cancer cells. The present study aimed to characterize ZFP91 expression in prostate cancer specimens. Furthermore, since our earlier reports showed discrepancies between ZFP91 mRNA and protein levels, we studied this interrelationship in LNCaP and PC-3 prostate cancer cell lines using siRNA mediated knockdown. QPCR analysis revealed marked upregulation of ZFP91 mRNA in the majority of prostate cancer specimens. Transfection of prostate cancer cells with ZFP91 siRNA resulted in a 10-fold decrease in mRNA levels. On a protein level, however, no inhibitory effect was observed over the time of the cell culture. We conclude that ZFP91 is overexpressed in prostate cancer and that potential accumulation of the ZFP91 protein in studied cells may be of importance in prostate cancer biology.

scholarly journals Effect of 1-Carbaldehyde-3,4-dimethoxyxanthone on Prostate and HPV-18 Positive Cervical Cancer Cell Lines and on Human THP-1 Macrophages

Molecules ◽  
2021 ◽  
Vol 26 (12) ◽  
pp. 3721
Author(s):  
Rui Medeiros ◽  
Bruno Horta ◽  
Joana Freitas-Silva ◽  
Jani Silva ◽  
Francisca Dias ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cervical Cancer ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Metabolic Activity ◽  
Cancer Cell Lines ◽  
No Production ◽  
Prostate Cell ◽  
Raw264.7 Macrophages

Xanthone derivatives have shown promising antitumor properties, and 1-carbaldehyde-3,4-dimethoxyxanthone (1) has recently emerged as a potent tumor cell growth inhibitor. In this study, its effect was evaluated (MTT viability assay) against a new panel of cancer cells, namely cervical cancer (HeLa), androgen-sensitive (LNCaP) and androgen-independent (PC-3) prostate cancer, and nonsolid tumor derived cancer (Jurkat) cell lines. The effect of xanthone 1 on macrophage functions was also evaluated. The effect of xanthone 1-conditioned THP-1 human macrophage supernatants on the metabolic viability of cervical and prostate cancer cell lines was determined along with its interference with cytokine expression characteristic of M1 profile (IL-1 ≤ β; TNF-α) or M2 profile (IL-10; TGF-β) (PCR and ELISA). Nitric oxide (NO) production by murine RAW264.7 macrophages was quantified by Griess reaction. Xanthone 1 (20 μM) strongly inhibited the metabolic activity of the cell lines and was significantly more active against prostate cell lines compared to HeLa (p < 0.05). Jurkat was the cell most sensitive to the effect of xanthone 1. Compound 1-conditioned IL-4-stimulated THP-1 macrophage supernatants significantly (p < 0.05) inhibited the metabolic activity of HeLa, LNCaP, and PC-3. Xanthone 1 did not significantly affect the expression of cytokines by THP-1 macrophages. The inhibiting effect of compound 1 observed on the production of NO by RAW 264.7 macrophages was moderate. In conclusion, 1-carbaldehyde-3,4-dimethoxyxanthone (1) decreases the metabolic activity of cancer cells and seems to be able to modulate macrophage functions.

scholarly journals Emblicanin-A Inhibits Cell Growth in Human Prostate Cancer Cells (PC-3) By Modulating Apoptotic Signaling Molecules

2020 ◽  
Vol 10 (5) ◽  
Author(s):  
Jayaraman Selvaraj ◽  
Rajagopal Ponnulakshmi ◽  
V Vishnupriya ◽  
Myneni Sindhura ◽  
Kamineni Sai Ravi Teja ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Mrna Expression ◽  
Cancer Cells ◽  
Caspase 3 ◽  
Treatment Options ◽  
B Cell Lymphoma ◽  
Multi Drug Resistance ◽  
Mrna Levels ◽  
Prostate Cancer Cells ◽  
Apoptotic Signaling

Cancer is a one of the leading causes of death in the world, continue to be worldwide dreadful disease. Prostate cancer is the most common cancer diagnosed in men. Multi-drug resistance (MDR) is a major problem with the current treatment options. It is now widely believed that many herbal dietary products are available as chemo-preventive agents against commonly occurring cancer types such as prostate cancer. Emblicanin-A, a hydrolyzable tannins has been isolated from the fruit of Emblicaofficinalis. The present study was aimedto find out whether emblicanin-A can inhibit growth of the prostate cancer cells (PC-3) through the regulation of apoptotic signaling mechanisms. Hence, PC-3 cells were treated with different concentrations of emblicanin-A (10, 25, 50, 100 and 150µM) for the analysis of B-cell lymphoma 2 (Bcl-2), Tumor suppressor protein (p53), Caspase-3 and caspase-9 mRNA expression in PC-3 cells. Cell viability was done using MTT in order to find the optimal dose. MTT assay exhibited that emblicanin-A showed cell death at the concentration of 100 and 150µM. It significantly (p<0.05) decreased the mRNA expression of anti apoptotic proteins (Bcl-2) while it upregulated the p53, caspase-3 and cas-9 mRNA levels effectively (p<0.05) in PC-3 cells which clearly indicates that emblicanin-A induces apoptosis in PC-3 cells by modulating intrinsic signalling mechanisms. To best of our knowledge, the present findings are the first to report anticancer activity of a bioactive compound from E.officinalis. Our study concludes that emblicanin-A may serve as a potential chemotherapeutic agent for the treatment of prostate cancer. Further studies on the effect of Emblicanin-A on the protein expression of further downstream signalling mechanisms is warranted in order to ascertain its potential mechanisms action towards clinical utility.

Deleted in cancer 1: Search for a function in prostate cancer

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. e16095-e16095
Author(s):  
J. Hirsch ◽  
T. Nelius ◽  
C. Pfarr ◽  
W. De Riese ◽  
I. Wieland ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Growth Factor ◽  
Cell Line ◽  
Cell Lines ◽  
Expression Profiling ◽  
Ectopic Expression ◽  
Mrna Levels ◽  
Normal Prostate ◽  
Prostate Cell ◽  
Growth Inhibitory

e16095 Background: Deleted In Cancer 1 (DICE1/INTS6) gene was recently identified to colocalize with the microsatellite marker D13S284 in 13q14.3, a region frequently affected by allelic deletion in many solid tumors including prostate cancer (PrCa). DICE1 missense mutations have been previously detected in PrCa cell line LNCap, and reduced DICE1 expression appears to be associated with CpG promoter hypermethylation in PrCa cells. DICE1 is a highly conserved nuclear protein suggesting its involvement in DNA repair, transcription, or RNA splicing. In mouse, the DICE1 homologue interferes with the response to insulin-like growth factor 1 and suppresses anchorage-independent growth of transformed mouse cells. The totality of these results suggests that DICE1 is a tumor suppressor gene and insist on the need to better characterize DICE1 function in PrCa. Methods: Expression of DICE1 was evaluated by Northern Blot in PrCa cell lines LNCap, DU145, PC3, PC3ml and CPTX1532 and compared to expression level in normal prostate cell line NPTX1532. DICE1 growth inhibitory effects were analyzed by colony formation assay on PC3 and DU145 cells transfected with DICE1 expression plasmid or control vector. Apoptosis was assessed by visualization of genomic DNA fragmentation on agarose gel. PCR arrays (SABiosciences) were used to identify specific signaling pathways modulated in response to DICE1 expression. Results: Markedly decreased DICE1 mRNA levels were detected in PrCa cell lines LNCap, DU145, PC3 and PC3ml as well as CPTX1532 as compared to NPTX1532, a cell line derived from normal prostate tissue. Ectopic expression of DICE1 cDNA in DU145 and PC3 cells substantially suppressed their ability to form colonies in vitro. This growth inhibition was not due to immediate induction of apoptosis suggesting growth suppression by other pathways. Expression profiling identified multiple pathways, such as Wnt, Hedgehog, PI-3 Kinase, NFκB and Insulin pathways, regulated in response to ectopic DICE1 expression. Furthermore, various transcription factors including Fos, Jun, CEBPA and PPAR-γ were up-regulated in response to DICE1. Conclusions: These results clearly illustrate the growth inhibitory ability of DICE1 in PrCa. Expression profiling links DICE1 function to growth factor signaling and cell-cell communication. No significant financial relationships to disclose.

scholarly journals Integrative proteomic and phosphoproteomic profiling of prostate cell lines

2019 ◽  
Author(s):  
Maria Katsogiannou ◽  
Jean-Baptiste Boyer ◽  
Alberto Valdeolivas ◽  
Elisabeth Remy ◽  
Laurence Calzone ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Line ◽  
Cell Lines ◽  
Differentially Expressed ◽  
Migration And Invasion ◽  
Mass Spectrometry Experiment ◽  
Disease Etiology ◽  
Prostate Cell ◽  
Functional Features ◽  
Prostate Cell Lines

ABSTRACTBackgroundProstate cancer is a major public health issue, mainly because patients relapse after androgen deprivation therapy. Proteomic strategies, aiming to reflect the functional activity of cells, are nowadays among the leading approaches to tackle the challenges not only of better diagnosis, but also of unraveling mechanistic details related to disease etiology and progression.MethodsWe conducted here a large SILAC-based Mass Spectrometry experiment to map the proteomes and phosphoproteomes of four widely used prostate cell lines, namely PNT1A, LNCaP, DU145 and PC3, representative of different cancerous and hormonal status.ResultsWe identified more than 3000 proteins and phosphosites, from which we quantified more than 1000 proteins and 500 phosphosites after stringent filtering. Extensive exploration of this proteomics and phosphoproteomics dataset allowed characterizing housekeeping as well as cell-line specific proteins, phosphosites and functional features of each cell line. In addition, by comparing the sensitive and resistant cell lines, we identified protein and phosphosites differentially expressed in the resistance context. Further data integration in a molecular network highlighted the differentially expressed pathways, in particular migration and invasion, RNA splicing, DNA damage repair response and transcription regulation.ConclusionsOverall, this study proposes a valuable resource toward the characterization of proteome and phosphoproteome of four widely used prostate cell lines and reveals candidates to be involved in prostate cancer progression for further experimental validation.

scholarly journals H3K27ac HiChIP in prostate cell lines identifies risk genes for prostate cancer susceptibility

2021 ◽  
Author(s):  
Claudia Giambartolomei ◽  
Ji-Heui Seo ◽  
Tommer Schwarz ◽  
Malika Kumar Freund ◽  
Ruth Dolly Johnson ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Lines ◽  
Cancer Susceptibility ◽  
Prostate Cell ◽  
Risk Genes ◽  
Prostate Cancer Susceptibility ◽  
Prostate Cell Lines

Proteomic analysis of laser capture microdissected human prostate cancer andin vitro prostate cell lines

2000 ◽  
Vol 21 (11) ◽  
pp. 2235-2242 ◽  
Author(s):  
David K. Ornstein ◽  
John W. Gillespie ◽  
Cloud P. Paweletz ◽  
Paul H. Duray ◽  
Judi Herring ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Lines ◽  
Proteomic Analysis ◽  
Human Prostate ◽  
Human Prostate Cancer ◽  
Prostate Cell ◽  
Laser Capture ◽  
Prostate Cell Lines ◽  
Laser Capture Microdissected
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