Although abundantly expressed in the nucleolus, Nucleolin also localizes to cytoplasmic and plasma membrane compartments [37C39], and its subcellular localization can indicate its functional role in different cell types [35]. to siCONT cells. Transfected HeLa cells were also assessed for changes in (C) migration and (D) invasion 6 h or 48 h following introduction of chemoattractant (FBS), respectively. siPVT1 cells showed a significant decrease in both cell migration and invasion compared to siCONT cells. Quantitative results are graphed on the left, while representative images are on the right. SC75741 **p<0.01, ***p<0.001, ****p<0.0001(TIF) pone.0156274.s003.tif (5.1M) GUID:?15E8C744-A6E3-479D-801B-3D52862BF6DE S4 Fig: PVT1 binds, but does not stabilize, Myc in SiHa cells. (A) PVT1 from SiHa total cell lysate immunoprecipitated with a MYC antibody, but not the negative control rabbit IgG (Cont). (B) PVT1 knockdown in SiHa cells did not significantly affect p58 phospho-MYC or total MYC protein. (C,D) Degradation of MYC protein was also not significantly affected by PVT1 knockdown. **p<0.01(TIF) pone.0156274.s004.tif (476K) GUID:?70EA7899-A6B4-4EAC-AC40-509BCE69A1CE S5 Fig: knockdown does not affect Nucleolin protein levels. (TIF) pone.0156274.s005.tif (971K) GUID:?84FAEDA5-E4E5-4D26-850B-5741EB99BB71 S1 Table: Percent knockdown of different regions achieved with siRNA or LNA. * denotes primer set used throughout the manuscript.(XLSX) pone.0156274.s006.xlsx (10K) GUID:?1FA64BD0-D3A8-45F1-A18E-BCA497DA3102 Data Availability StatementAll relevant data are within the paper and its Supporting Information files. Abstract The plasmacytoma variant translocation 1 gene (influences disease processes has been studied in multiple cancer types, its role in cervical tumorigenesis remains unknown. Thus, the present study was designed to investigate the role of in cervical cancer and expression was measured by quantitative PCR (qPCR) in 121 invasive cervical carcinoma (ICC) samples, 30 normal cervix samples, and cervical cell lines. Functional assays were carried out using both siRNA and LNA-mediated knockdown to examine expression is SC75741 significantly increased in ICC tissue versus normal cervix and that higher expression of correlates with poorer overall survival. In cervical cancer cell lines, knockdown resulted in significantly decreased cell proliferation, migration and invasion, while apoptosis and cisplatin cytotoxicity were significantly increased in these cells. Finally, we show that expression is augmented in response to hypoxia and immune response stimulation and that this lncRNA associates with the multifunctional and stress-responsive protein, Nucleolin. Collectively, our results SC75741 provide strong evidence for an oncogenic role of in cervical cancer and lend insight into potential mechanisms by which overexpression helps drive cervical carcinogenesis. Introduction Long noncoding RNAs (lncRNAs) provide important targets for cancer diagnostics and therapeutics due to their critical role in numerous cellular processes such as epigenetic changes, gene enhancer and tumor suppressor activity, and miRNA sequestering. LncRNAs are pervasive in the genome, frequently show cell type- and temporal-specific regulation of gene expression, and can influence Mouse monoclonal antibody to AMPK alpha 1. The protein encoded by this gene belongs to the ser/thr protein kinase family. It is the catalyticsubunit of the 5-prime-AMP-activated protein kinase (AMPK). AMPK is a cellular energy sensorconserved in all eukaryotic cells. The kinase activity of AMPK is activated by the stimuli thatincrease the cellular AMP/ATP ratio. AMPK regulates the activities of a number of key metabolicenzymes through phosphorylation. It protects cells from stresses that cause ATP depletion byswitching off ATP-consuming biosynthetic pathways. Alternatively spliced transcript variantsencoding distinct isoforms have been observed many cellular processes via multiple disparate mechanisms [1]. Plasmacytoma variant translocation 1 (that has attracted significant attention from the cancer field due to its frequent co-amplification with in several solid tumors [2]. The first studies providing evidence that may contribute to carcinogenesis demonstrated frequent translocations in mouse plasmacytomas [3,4] and human Burkitts lymphomas [5C7]. The oncogenic effects of have been further highlighted by more recent studies demonstrating its overexpression and amplification in multiple cancer types [8C17]. More importantly, expression has been significantly correlated with clinical features such as risk, recurrence, and survival in various cancers [8,11C13,18]. Despite the wealth of knowledge regarding the oncogenic properties of in multiple cancers, very little is known about its precise biologic function. In fact, the handful of studies providing mechanistic data exceedingly suggest that it exerts its effects in a cell-type and/or disease-specific manner. For example, function has been attributed to its binding and stabilization of the Myc [19] and Nop2 [17] proteins in breast and hepatocellular carcinoma, respectively. In gastric cancer cells, acts to repress the expression of p15 and p16 via its physical interaction with the polycomb group protein, EZH2 [20]. Also via EZH2 recruitment and regulation of thyroid-stimulating hormone receptor, induces proliferation of thyroid cancer cells [21]. Finally, computational analysis of suggests that it may act via binding and sequestration of mir-200 family members in breast cancer tissue [14]. Due to their complexity and breadth of results, these studies emphasize the importance of disease-specific investigation of mechanisms. Our interest in originated from our.