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LINC01094 promotes the invasion of ovarian cancer cells and regulates the Wnt/β-catenin signaling pathway by targeting miR-532-3p.

ABSTRACT: Long non-coding RNAs (lncRNAs) participate in the development of ovarian cancer (OC). The present study aimed to explore the roles of long intergenic non-protein coding RNA 1094 (LINC01094) in OC. LINC01094 and microRNA (miR)-532-3p expression in OC tissues and cells were measured using reverse transcription-quantitative PCR. Cell migration and invasion were detected using wound healing assays and Transwell assays, respectively. The binding of LINC01094 or β-catenin to miR-126-5p was detected using a Dual-luciferase reporter assay, and protein expression was confirmed using western blot analysis. The expression level of LINC01094 in patients with OC was higher in OC tissues compared with in adjacent tissues, and LINC01094 was upregulated in OC cell lines. In addition, LINC01094 overexpression promoted the viability, migration, invasion and cell cycle progression of OC cells, and inhibited OC cell apoptosis. Moreover, LINC01094 negatively regulated miR-532-3p in OC cells and tissues. miR-532-3p overexpression decreased the viability, migration, invasion and cell cycle progression of OC cells alongside downregulation of Wnt/β-catenin signaling pathway protein expression, as well as increasing OC cell apoptosis. Inhibition of LINC01094 with small interfering (si)-LINC01094 and overexpression of LINC01094 respectively reversed the effect of miR-532-3p inhibitor and mimics on OC cells. miR-532-3p could directly target β-catenin, and miR-532-3p inhibitor increased β-catenin expression, while si-LINC01094 attenuated this effect. In addition, LINC01094 overexpression promoted tumor growth in vivo by regulating miR-532-3p. Taken together, LINC01094 promoted the growth, migration, invasion and Wnt/β-catenin signaling pathway expression of OC cells by modulating miR-532-3p.

SUBMITTER: Chen H

PROVIDER: S-EPMC8438678 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BackgroundEsophageal squamous cell carcinoma (ESCC) is the predominant subtype of esophageal cancer in East Asia, with approximately half of ESCC cases occurring in China. ESCC poses a serious threat to the quality of life of patients. MicroRNAs (miRNAs) are extremely important in the occurrence and development of ESCC. Current studies have shown that miR-212-3p is expressed at low levels in esophageal adenocarcinoma tumor tissues; however, its function and mechanism in ESCC have not been studied.MethodsThe expression levels of miR-212-3p and SOX4 were detected by quantitative polymerase chain reaction (qPCR) in ESCC tissues and adjacent tissues, and ESCC cell lines from 60 patients. The interaction of miR-212-3p and SOX4 was determined using a dual-luciferase reporter gene system. EC9706 Cells were transfected with miR-212-3p mimic, NC mimic, si-NC and si-SOX4 and miR-212-3p mimic + overexpressing-SOX4, and the effects on miR-212-3p and SOX4 expression were observed, respectively. An MTT assay was carried out to detect the proliferation ability of ESCC. The invasion ability and apoptosis level of the cells were determined by Transwell assay and flow cytometry, respectively. qPCR was used to detect expression of miR-212-3p and SOX4. Western blot was performed to observe the expression of SOX4, Wnt1, β-catenin, c-Myc, and Cyclin D1.ResultsmiR-212-3P was observed to be down-regulated in ESCC tissues and cells, while SOX4 expression was up-regulated; the two were negatively correlated. The dual-luciferase reporter gene further confirmed that miR-212-3p targeted SOX4. miR-212-3p overexpression and interference with SOX4 significantly inhibited the proliferation and invasion of ESCC EC970 cells, and promoted apoptosis. Furthermore, the results of Western blot confirmed that miR-212-3p overexpression and interference with SOX4 down-regulated the expression of Wnt1, β-catenin, c-Myc, and Cyclin D1. Meanwhile, SOX4 overexpression reversed the effect of up-regulation of miR-212-3p on EC970 function.ConclusionsmiR-212-3p mediates the apoptosis and invasion of ESCC cells through inhibiting the Wnt/β-catenin signal pathway by targeting SOX4.

Project description:ObjectiveThis study aimed to investigate the effect of Wnt/?-catenin signal pathway mediated by miR-342-5p targeting CBX2 gene on the proliferation, metastasis, invasion and apoptosis of ovarian cancer cells, and to explore its related regulatory mechanism.MethodsHuman normal ovarian epithelial cell line IOSE80, human ovarian cancer cell line SKOV3 and OVCAR3 were the subjects. Software were used to predict the binding site of miR-342-5p targeting CBX2 gene. The proliferation rate of ovarian cancer cells was detected by MTT method; the cell viability of each group was observed by colony formation test; the apoptosis of cells in each group was detected by flow cytometry; the invasive ability of cells was determined by transwell test, and the migration ability of cells was detected by scratch test. The mRNA expression levels of miR-342-5p, CBX2, Wnt1, ?-catenin, C-myc and Cyclin D1 were measured by qRT-PCR. Also, Western blot was used to determine the protein expression levels of CBX2, Wnt1, ?-catenin, C-myc and Cyclin D1.ResultsCBX2 was identified as the target gene of miR-342-5p. MTT test results showed that miR-342-5p could significantly inhibit the proliferation of SKOV3 and OVCAR3 cells, colony formation assay results indicated that the viability of SKOV3 and OVCAR3 cells transfected with miR-342-5p decreased significantly, and flow cytometry results suggested that miR-342-5p could promote the apoptosis of SKOV3 and OVCAR3 cells. Also, the results of transwell showed that miR-342-5p could significantly inhibit the invasive ability of SKOV3 and OVCAR3 cells, and the results of scratch assay suggested that miR-342-5p could significantly inhibit the migration of SKOV3 and OVCAR3 cells. Moreover, qRT-PCR and Western blot results indicated that the mRNA and protein expression levels of CBX2, Wnt1, ?-catenin, C-myc and Cyclin D1 decreased in SKOV3 and OVCAR3 cells transfected with miR-342-5p, while the mRNA expression levels of miR-342-5p increased significantly (P<0.05).ConclusionMiR-342-5p targeted gene is CBX2, which can significantly reduce the proliferation, invasion, migration and viability of ovarian cancer cell lines SKOV3 and OVCAR3, and promote their apoptosis. The mechanism may be related to the mediation of Wnt/?-catenin signal pathway and down-regulation of the related genes expression.

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LINC01094 promotes the invasion of ovarian cancer cells and regulates the Wnt/β-catenin signaling pathway by targeting miR-532-3p.

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