UNASSIGNED:本研究旨在研究circ_0000799对结直肠癌(CRC)细胞生物学功能的影响及其机制。
未经批准:首先,定量逆转录聚合酶链反应(qRT-PCR)用于检测circ_0000799,miR-647和miR-1243在住院CRC患者手术切除标本中的表达,CRC-癌旁正常组织(正常组),人正常结肠上皮细胞(FHC组),和CRC细胞系(HCT116,HT29,SW480,SW620)。细胞增殖,生存能力,并通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化物(MTT)检测到侵袭,集落形成试验,在过表达或抑制circ_0000799的HCT116和SW480细胞中进行transwell测定。circ_0000799和miR-647之间的靶向关系通过双荧光素酶报告基因实验进行验证。上皮间质转化(EMT)蛋白(E-cadherin,波形蛋白,和N-钙黏着蛋白)通过蛋白质印迹进行测试。
UNASSIGNED:CRC组织和细胞中circ_0000799的表达水平显着增加。过表达circ_0000799显著提高细胞增殖率,生存能力,入侵,和EMT过程,而敲除circ_0000799抑制CRC细胞的生物学性能。生物信息学分析提示miR-647受circ_0000799调控,双荧光素酶报告基因检测进一步显示了二者之间的靶向关系。此外,circ_0000799与miR-647表达呈负相关。
UNASSIGNED:我们的研究结果表明circ_0000799促进CRC和EMT的增殖和侵袭。circ_0000799的这些作用可以通过负调控miR-62来实现。
UNASSIGNED: The current study aimed to investigate the effect of circ_0000799 on the biological function of colorectal cancer (CRC) cells and its mechanism.
UNASSIGNED: First, quantitative reverse transcription polymerase chain reaction (qRT-PCR) was employed for detecting the expression of circ_0000799, miR-647, and miR-1243 in surgically resected specimens from hospitalized CRC patients, CRC-adjacent normal tissues (Normal group), human normal colon epithelial cells (FHC group), and CRC cell lines (HCT116, HT29, SW480, SW620). The cell proliferation, viability, and invasion were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), colony formation assay, transwell assay in HCT116 and SW480 cells with overexpression or inhibition of circ_0000799. The targeting relationship between circ_0000799 and miR-647 was verified by dual-luciferase reporter assay. The expression of epithelial-mesenchymal transition (EMT) proteins (E-cadherin, vimentin, and N-cadherin) was tested by western blot.
UNASSIGNED: The expression level of circ_0000799 was significantly increased in CRC tissues and cells. Overexpression of circ_0000799 significantly increased cell proliferation rate, viability, invasion, and the EMT process, whereas knockdown of circ_0000799 inhibited the biological performance of CRC cells. Bioinformatic analysis suggested that miR-647 was regulated by circ_0000799, and a dual-luciferase reporter assay further showed a targeting relationship between the two. In addition, circ_0000799 was negatively correlated with miR-647 expression in CRC.
UNASSIGNED: Our findings suggest that circ_0000799 promotes proliferation and invasion in CRC and EMT. These effects of circ_0000799 may be achieved by negatively regulating miR-62.