Long noncoding RNA ZFAS1 promotes gastric cancer cells proliferation by epigenetically repressing KLF2 and NKD2 expression.

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In this study, we globally assessed the transcriptomic differences of lncRNAs in gastric cancer using publicly available microarray data from Gene Expression Omnibus (GEO) and identified an oncogenetic lncRNA ZFAS1, which may promote gastric tumorigenesis. ZFAS1 has been found to be upregulated and function as oncogene in colorectal cancer and hepatocellular carcinoma, but its expression pattern, biological function and underlying mechanism in gastric cancer is still undetermined. Here, we reported that ZFAS1 expression is also overexpressed in gastric cancer, and its increased level is associated with poor prognosis and shorter survival. Knockdown of ZFAS1 impaired gastric cancer cells proliferation and induced apoptosis in vitro, and inhibited tumorigenicity of gastric cancer cells in vivo. Mechanistically, RNA immunoprecipitation and RNA pull-down experiment showed that ZFAS1 could simultaneously interact with EZH2 and LSD1/CoREST to repress underlying targets KLF2 and NKD2 transcription. In addition, rescue experiments determined that ZFAS1 oncogenic function is partly dependent on repressing KLF2 and NKD2. Taken together, our findings illuminate how ZFAS1 over-expression confers an oncogenic function in gastric cancer.", "claims": [{"id": "claim-method", "statement": "The Students t test (2 tailed), one-way ANOVA, and Mann-Whitney U test were conducted to analyze the in vitro and in vivo data by SPSS 17.0 software.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:27246976", "locator": "sections[25] \"Statistical analysis\" paragraph 2", "quote": "The Students t test (2 tailed), one-way ANOVA, and Mann-Whitney U test were conducted to analyze the in vitro and in vivo data by SPSS 17.0 software."}], "state": "complete"}, {"id": "claim-data-access", "statement": "Four human microarray datasets including GSE37023, GSE13911, GSE65801 and GSE51575 were obtained from public GEO database (http://www.ncbi.nlm.nih.gov/geo) and normalized using Robust Multichip Average (RMA).", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:27246976", "locator": "sections[13] \"Microarray data analysis\" paragraph 2", "quote": "Four human microarray datasets including GSE37023, GSE13911, GSE65801 and GSE51575 were obtained from public GEO database (http://www.ncbi.nlm.nih.gov/geo) and normalized using Robust Multichip Average (RMA)."}], "state": "complete"}, {"id": "claim-target-result", "statement": "The paper’s headline result is that ZFAS1 is overexpressed in gastric cancer and its increased level is associated with poor prognosis and shorter survival: “Here, we reported that ZFAS1 expression is also overexpressed in gastric cancer, and its increased level is associated with poor prognosis and shorter survival.” No numerical result is stated for the registered target, which is a statistical method and data route rather than a numerical outcome.", "kind": "qualitative", "target": null, "evidence": [{"asset": "source:pmid:27246976", "locator": "sections[0] \"Abstract\" paragraph 1", "quote": "Here, we reported that ZFAS1 expression is also overexpressed in gastric cancer, and its increased level is associated with poor prognosis and shorter survival."}, {"asset": "source:pmid:27246976", "locator": "sections[25] \"Statistical analysis\" paragraph 2", "quote": "The Students t test (2 tailed), one-way ANOVA, and Mann-Whitney U test were conducted to analyze the in vitro and in vivo data by SPSS 17.0 software. P values less than 0.05 were considered significant."}, {"asset": "source:pmid:27246976", "locator": "sections[13] \"Microarray data analysis\" paragraph 2", "quote": "Four human microarray datasets including GSE37023, GSE13911, GSE65801 and GSE51575 were obtained from public GEO database (http://www.ncbi.nlm.nih.gov/geo) and normalized using Robust Multichip Average (RMA)."}, {"asset": "source:pmid:27246976", "locator": "sections[3] \"lncRNA ZFAS1 is overexpressed in gastric cancer tissues and cell lines\" paragraph 2", "quote": "To identify lncRNAs involved in gastric cancer development and progression, four microarray datasets (GSE37023, GSE13911,GSE65801 and GSE51575) were obtained from GEO to analyze lncRNAs alterations between gastric cancer and pair nontumor tissues."}, {"asset": "source:pmid:27246976", "locator": "sections[3] \"lncRNA ZFAS1 is overexpressed in gastric cancer tissues and cell lines\" paragraph 2", "quote": "In addition, analysis of TCGA stomach and normal tissues RNA sequencing data also showed that ZFAS1 expression is up-regulated in tumor tissues compared with normal tissues (Figure 1C)."}], "state": "complete"}], "gaps": []}, "experiment": {"state": "partial", "objective": "The paper’s headline result is that ZFAS1 is overexpressed in gastric cancer and its increased level is associated with poor prognosis and shorter survival: “Here, we reported that ZFAS1 expression is also overexpressed in gastric cancer, and its increased level is associated with poor prognosis and shorter survival.” No numerical result is stated for the registered target, which is a statistical method and data route rather than a numerical outcome.", "steps": [{"id": "read_published_estimate", "order": 1, "name": "Read the published estimate from the source", "kind": "other", "state": "complete", "blocked_on": null, "description": "The claim, method and evidence span published with the source article.", "duration_hours": null, "parameters": {}, "inputs": ["source:pmid:27246976"], "outputs": ["the published estimate and its evidence"], "instruments": [], "reagents": []}, {"id": "obtain_source_data", "order": 2, "name": "Obtain the source data from the named repository", "kind": "data-preparation", "state": "complete", "blocked_on": null, "description": "The GEO series accession resolves and its sample accessions are listed. 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