The long non-coding RNA LINC00152 is essential for cell cycle progression through mitosis in HeLa cells

Abstract

In recent years, long non-coding RNA (lncRNA) research has identified essential roles of these transcripts in virtually all physiological cellular processes including tumorigenesis, but their functions and molecular mechanisms are poorly understood. In this study, we performed a high-throughput siRNA screen targeting 638 lncRNAs deregulated in cancer entities to analyse their impact on cell division by using time-lapse microscopy. We identified 26 lncRNAs affecting cell morphology and cell cycle including LINC00152. This transcript was ubiquitously expressed in many human cell lines and its RNA levels were significantly upregulated in lung, liver and breast cancer tissues. A comprehensive sequence analysis of LINC00152 revealed a highly similar paralog annotated as MIR4435-2HG and several splice variants of both transcripts. The shortest and most abundant isoform preferentially localized to the cytoplasm. Cells depleted of LINC00152 arrested in prometaphase of mitosis and showed reduced cell viability. In RNA affinity purification (RAP) studies, LINC00152 interacted with a network of proteins that were associated with M phase of the cell cycle. In summary, we provide new insights into the properties and biological function of LINC00152 suggesting that this transcript is crucial for cell cycle progression through mitosis and thus, could act as a non-coding oncogene.

The Long Noncoding RNA Cancer Susceptibility 9 and RNA Binding Protein Heterogeneous Nuclear Ribonucleoprotein L Form a Complex and Coregulate Genes Linked to AKT Signaling

“…We applied functional siRNA screening for lncRNAs with a loss‐of‐viability phenotype in HCC. The results of the screening are in excellent accordance with other studies: the lncRNA genes HOX transcript antisense RNA, long intergenic non–protein coding RNA 152, Pvt1 oncogene, and HCC up‐regulated lncRNA are known to be important for cancer cell viability and exhibited high negative z scores (loss of viability) in the screen. Hence, this data set provides a useful resource for subsequent research on viability‐associated lncRNAs in HCC.…”

“…To derive a broader set of cancer‐associated lncRNAs and to design a library of siRNAs against these candidates as a versatile tool to dissect lncRNAs in cancer, this data set was merged with results from two other cancer entities, breast ductal carcinoma and lung adenocarcinoma (all data deposited in the National Center for Biotechnology Information’s Gene Expression Omnibus). The analysis of this integrated data set yielded a total of 638 significantly dysregulated lncRNAs, mostly up‐regulated, which were used for the siRNA library design as described . An RNAi‐mediated knockdown screen of these 638 candidate genes was done, and the impact on cellular viability in two established HCC cell lines (HLE and Huh‐7) was determined.…”

“…The small interfering RNA (siRNA) library, consisting of >3,100 siRNAs targeting 638 potentially oncogenic lncRNAs, was designed as described . Reverse transfection was performed in 384 well plates with 30 nM siRNA in a volume of 50 μL (5 μL siRNA in H 2 O, 0.05 μL RNAiMAX; Thermo Fisher Scientific), 14.95 μL Roswell Park Memorial Institute medium, and 30 μL cells in growth medium.…”

Improved characterization of the relationship between long intergenic non‐coding RNA Linc00152 and the occurrence and development of malignancies

“…They also demonstrated that the knockdown of linc00152 by a prometaphase arrest can indirectly induce the apoptosis of transfected HeLa cells, further implying that linc00152 may participate in the cell cycle by interacting with proteins related to the M phase; however, the mechanism mediating this interaction remains unclear. 54 Other studies have also suggested that linc00152 affects the cell cycle. 27,29,34,36,54…”

“…Nötzold et al detected upregulated linc00152 expression in lung, liver, and breast cancers, but not in Burkitt's lymphoma. A cell viability assay indicated that inhibited linc00152 expression can disrupt cell proliferation.…”