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Long non-coding RNA LINC00460 promotes proliferation and inhibits apoptosis of cervical cancer cells by targeting microRNA-503-5p.
Molecular and Cellular Biochemistry ( IF 3.5 ) Pub Date : 2020-08-01 , DOI: 10.1007/s11010-020-03853-0
Lin Lin 1 , Bing Xin 2 , Tao Jiang 2 , Xin-Lu Wang 1 , Hua Yang 1 , Tie-Mei Shi 1
Affiliation  

Long non-coding RNAs are associated with the pathogenesis of cancers. Moreover, LINC00460 is involved in the development of multiple cancers. However, the function of LINC00460 in cervical cancer (CC) remains inconclusive. Herein, CC tissues and tumor-adjacent tissues were collected from patients. The effect of LINC00460 silencing in cell proliferation and apoptosis in CC was explored in vitro and in vivo. Additionally, the interaction between LINC00460 and miR-503-5p was analyzed using dual luciferase reporter assay. The expression of genes and proteins was assayed using quantitative real-time PCR, western blotting and immunohistochemistry, cell viability using MTT assay, cell cycle distribution using flow cytometry, cell apoptosis using Annexin V staining, Hoechst staining and TUNEL assay. LINC00460 levels in CC tissues were higher than tumor-adjacent tissues. LINC00460 silencing suppressed proliferation and promoted apoptosis of CC cells as evidenced by decreased cell viability, inhibited proliferation-related protein and cell cycle protein expressions and G1/S transition, increased apoptotic cells and Hoechst-positive cells, and enhanced apoptosis-related protein expressions. LINC00460 could bind to miR-503-5p and LINC00460 silencing enhanced miR-503-5p expression and inhibited its target gene expressions in CC cells. MiR-503-5p inhibition reversed LINC00460 silencing-caused inhibition of cell proliferation and miR-503-5p target gene expressions, and promotion of cell apoptosis. LINC00460 silencing also attenuated tumor growth, promoted miR-503-5p levels and cell apoptosis, and inhibited cell proliferation and miR-503-5p target gene expressions in tumor tissues. Hence, LINC00460 functioned as an oncogene in CC that affected cell proliferation and apoptosis via sponging miR-503-5p. This study provides a novel therapeutic target for CC.



中文翻译:

长的非编码RNA LINC00460通过靶向microRNA-503-5p促进宫颈癌细胞的增殖并抑制其凋亡。

长的非编码RNA与癌症的发病机制有关。此外,LINC00460参与了多种癌症的发展。但是,LINC00460在子宫颈癌(CC)中的功能仍不确定。在此,从患者收集CC组织和肿瘤附近组织。体外和体内研究了LINC00460沉默对CC细胞增殖和凋亡的影响。另外,使用双重荧光素酶报告基因分析法分析了LINC00460和miR-503-5p之间的相互作用。使用定量实时PCR,蛋白质印迹和免疫组织化学,使用MTT分析的细胞活力,使用流式细胞术的细胞周期分布,使用Annexin V染色,Hoechst染色和TUNEL分析的细胞凋亡来检测基因和蛋白质的表达。CC组织中的LINC00460水平高于肿瘤邻近组织。LINC00460沉默可抑制CC细胞增殖,并促进CC细胞凋亡,其表现为细胞活力降低,抑制增殖相关蛋白和细胞周期蛋白表达以及G1 / S过渡,凋亡细胞和Hoechst阳性细胞增加以及凋亡相关蛋白表达增强。LINC00460可以与miR-503-5p结合,而LINC00460沉默增强的miR-503-5p表达,并抑制CC细胞中其靶基因的表达。MiR-503-5p抑制可逆转LINC00460沉默引起的细胞增殖和miR-503-5p靶基因表达的抑制,并促进细胞凋亡。LINC00460沉默还可减缓肿瘤生长,促进miR-503-5p水平和细胞凋亡,并抑制肿瘤组织中的细胞增殖和miR-503-5p靶基因表达。因此,LINC00460在CC中起着癌基因的作用,通过海绵化miR-503-5p影响细胞的增殖和凋亡。该研究为CC提供了新的治疗靶标。

更新日期:2020-08-01
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