Abstract
BackgroundThe study purpose was to make investigation into the influence of XIST on cervical cancer progression and what’s more its potential mechanism.MethodsThe cervical cancer data sets (lncRNA, miRNA, and mRNA) obtained from TCGA were analyzed with the “mixOmics” R package. Then, the expression of XIST, miR-140-5p, and ORC1 were detected using qRT-PCR and western blot in both tissues and cervical cancer cell lines (Hela and C33A) to verify the bioinformatics analyses results. CCK-8 assay, 5-ethynyl-2′-deoxyuridine (EdU) assays, cell cycle assay and cell apoptosis assay were practiced. Besides, immunohistochemistry staining was operated for the detection of the Ki-67, E-cadherin and vimentin expression in cervical cancer tissues and the apoptosis-related proteins expression (c-caspase3, Bcl-2, total PARP and cleaved PARP) was verified through western blot. And in vivo experiments were implemented.ResultsMiR-140-5p was down-regulated but XIST and ORC1 were up-regulated in cervical cancer tissues and cell lines. Knocking down of the XIST or ORC1 memorably suppressed cell proliferation, blocked cell cycle, decreased the expression of Bcl-2 while increased the apoptosis rate and the expression of c-caspase3 and cleaved PARP in HeLa and C33A cells. Besides, the results of immunohistochemistry staining showed knocking down the expression of XIST improved the expression levels of E-cadherin and decreased Ki-67 and vimentin expression. And overexpression of miR-140-5p also could inhibit the progression and reverse the influence of XIST and ORC1 in HeLa and C33A cells.ConclusionOur study indicated the effects of XIST/miR-140-5p/ORC1 axis on the progression of cervical cancer which will shed new light on epigenetic diagnostics and therapeutics in cervical cancer.
Highlights
The study purpose was to make investigation into the influence of X-inactive specific transcript (XIST) on cervical cancer progression and what’s more its potential mechanism
The clustered image map (CIM) on the basis of the miRNA, messenger RNAs (mRNA) and long non-coding RNAs (lncRNAs) selected on the first component well classified the tumor samples as well as normal samples (Fig. 2a)
Results showed that XIST and ORC1 expression was significantly down-regulated in si-XIST injection group compared to control group after 7 weeks, while miR-140-5p expression was conversed (Fig. 4h, i). These results suggested that inhibition of XIST could significantly suppress proliferation capacity of cervical cancer in vivo, and these effects might be achieved through miR-140-5p and ORC1
Summary
The study purpose was to make investigation into the influence of XIST on cervical cancer progression and what’s more its potential mechanism. After the targeting interaction between long non-coding RNAs (lncRNA), messenger RNAs (mRNA) and microRNA (miRNA) being found, more biological medicine researches and treatments turns to gene therapy field, and many investigations related with the regulations between various types of RNAs and proteins in cancer cells have been undertaken. LncRNA targeting miRNA and mRNA in cervical cancer cells. MixOmics analysis could be applied in exploring the biological data set by means of multivariate analysis, evaluating the molecular interactions happened at multiple levels in function and acquiring a large scale of information simultaneously [5]. We used mixOmics to integrate different data sets and attempted to reveal the potential molecular mechanism of cervical cancer
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