Mechanisms Of LncRNA-UCA1 On Gastrle Cancer Progesslon

ObjectiveLong non-coding RNAs(lncRNAs) is a class of non-protein-coding genomic transcripts. lncRNAs influence the fundamental biological processes and the development of diseases including cancers through a variety of regulatory mechanisms. In this study, gastric cancer related lncRNAs were screened by analyzing transcriptome microarray data and RNAseq data. We found that the expression of lncRNA-urothelial cancer associated 1(UCA1) was increased in gastric cancer, and then explored the mechanisms underlying its impacts on gastric cancer development.Methods1. The expression of UCA1 in gastric cancer tissues was examined by multiple sets of transcriptome microarray data and RNAseq data. Copy number variation of UCA1 gene in gastric cancer was analyzed by online tool cBioPortal. The correlation between UCA1 expression levels and clinicopathological characteristics of gastric cancer was analyzed by The Cancer Genome Atlas(TCGA) data.2. The relative expression level of UCA1 in gastric epithelial cell line and gastric cancer cell lines was detected by real-time quantitative reverse transcrition-polymerase chain reaction(qRT-PCR). Gastric cancer cell line AGS was selected as the model cell line. RNA-FISH assay was used to analyze the subcellular location of UCA1 in gastric cancer cells.3. Following knock-down of UCA1 by siRNA transfection, the cell proliferation was investigated by MTT assay; and the cell cycle and apoptosis was analyzed by flow cytometry.4. After silencing of UCA1 by lentiviral-mediated shRNA, cell proliferation ability was performed by cell count, high content analysis and colony formation assays; cell migration and metastasis ability was assayed by high content analysis, wound healing assays and Transwell assay, respectively.5. Screening differentially expressed mi RNA in gastric cancer by TCGA mi Rseq data and predicting miRNA binding to UCA1 were combined to explore potential interactions between UCA1 and miRNA.6. The proteins regulated by UCA1 were analyzed by Mass Spectrometry.Results1. Analysis of transcriptome microarray data and RNAseq data indicated that UCA1 was up-regulated in gastric cancer. However, its expression level does not have any relationship with its genomic copy number variation and clinicopathological characteristics of patients with gastric cancer analyzed by TCGA data.2. Compared with normal gastric epithelial cell line GES, UCA1 expression level in AGS cells was significantly higher. RNA-FISH assay showed that UCA1 mainly located in the cytoplasm.3. Silencing of UCA1 by siRNA significantly inhibited gastric cancer cell proliferation and arrested cell cycle at G1 to S transition, but did not induce cell apoptosis. We found that UCA1 regulated the distributions of cell cycle and affected gastric cancer cell proliferation by regulating cyclin-dependent kinase inhibitor 1B(CDKN1B) and cell division cycle 25B(CDC25B).4. Knock-down of UCA1 by shRNA inhibited gastric cancer cell proliferation, migration and metastasis.5. Data analysis indicated that mi R-143 might interact with UCA1 and knock-down of UCA1 reduced the mRNA expression level of hexokinase(HK2), a verified target of phenotypes in gastric cancer cells. We found that UCA1 regulated the distributions of cell cycle and affected gastric cancer cell proliferation by regulating CDKN1 B and CDC25 B and may affect glucose metabolism through combining with miR-143 to regulate HK2 expression in gastric cancer cells. In addition, it also regulated the protein expression of TP53 and p-AKT to influence cell proliferation and metastasis of gastric cancer. Taken together, the present study provides a potential target for the treatment of gastric cancer.miR-143. 6. The result of Mass Spectrometry showed that knock-down of UCA1 affected tumor-associated pathways, such as, cell cycle, cell adhesion and TP53 signaling pathway, etc. ConclusionsBased on analysis of gastric cancer transcriptome microarray data and RNAseq data, we identified the lnc RNA-UCA1 which associated with gastric cancer cell proliferation, migration and metastasis and explore its underlying mechanism contributing to a variety of phenotypes in gastric cancer cells. We found that UCA1 regulated the distributions of cell cycle and affected gastric cancer cell proliferation by regulating CDKN1 B and CDC25 B and may affect glucose metabolism through combining with mi R-143 to regulate HK2 expression in gastric cancer cells. In addition, it also regulated the protein expression of TP53 and p-AKT to influence cell proliferation and metastasis of gastric cancer. Taken together, the present study provides a potential target for the treatment of gastric cancer.