环状RNA BCRC-3对鼻咽癌细胞增殖和迁移的影响研究

Effect of circular RNA BCRC-3 on the proliferation and migration of nasopharyngeal carcinoma cells

目的观察鼻咽癌组织和细胞株中环状RNA(circular RNA,circRNA)BCRC-3的表达,探讨环状RNA BCRC-3对细胞周期依赖性激酶抑制蛋白B(cyclin dependent kinase inhibitors B,DKNIB)基因表达的调控及对鼻咽癌细胞增殖和迁移的影响。方法荧光实时定量聚合酶链式反应(qPCR)分别检测circRNA BCRC-3在83例鼻咽癌组织及62例慢性鼻咽炎组织、鼻咽癌细胞株及人永生化鼻咽上皮细胞中的表达。以circRNA BCRC-3表达最低的细胞株为转染对象,分别转染阴性对照质粒(对照组)或载有circRNA BCRC-3序列的质粒(实验组)。MTS法和细胞划痕实验分别检测上调circRNA BCRC-3对细胞增殖和迁移能力的影响。qPCR检测上调环状RNA BCRC-3对DKNIB基因mRNA表达的影响,Western blot检测DKNIB蛋白和PI3K/AKT信号通路蛋白的表达。结果与慢性鼻咽炎组织相比,circRNA BCRC-3在鼻咽癌组织中表达显著降低(P<0.01)。与人永生化鼻咽上皮细胞相比,环状RNA BCRC-3在鼻咽癌细胞株中表达显著降低(P<0.01),在HONE-1细胞中的表达最少(P<0.01)。与对照组比较,实验组HONE-1细胞增殖能力从第2天开始明显下降(P<0.05),HONE-1细胞迁移能力明显降低(P<0.01),HONE-1细胞中DKNIB在mRNA和蛋白水平的表达明显上升(P<0.01),PI3K/AKT信号通路蛋白PIP3、PDK1、p-AKT和AS160表达明显下降。结论circRNA BCRC-3在鼻咽癌组织和细胞株中呈低表达,上调circRNA BCRC-3可抑制鼻咽癌HONE-1细胞的增殖和迁移能力,其机制可能是circRNA BCRC-3通过上调DKNIB基因表达,抑制PI3K/AKT信号通路转导。

OBJECTIVE To observe the expression of circular RNA(circRNA)BCRC-3 in nasopharyngeal carcinoma tissues and cell lines,and to explore the regulation of circRNA BCRC-3 on cyclin dependent kinase inhibitors B(DKNIB)gene expression and its effect on the proliferation and migration of nasopharyngeal carcinoma cells.METHODS Fluorescent real-time quantitative polymerase chain reaction(qPCR)was used to detect the expression of circRNA BCRC-3 in 83 cases of nasopharyngeal carcinoma tissue,62 cases of chronic nasopharyngitis tissue,nasopharyngeal carcinoma cell line,and human immortalized nasopharyngeal epithelial cells.The cell line with the lowest circRNA BCRC-3 expression was used as the transfection cells,and the negative control plasmid(control group)or the plasmid containing the circRNA BCRC-3 sequence(experimental group)was transfected.MTS method and Transwell migration assay were used to detect the effects of up-regulated circRNA BCRC-3 on cell proliferation and migration ability.qPCR was used to detect the effect of up-regulated circRNA BCRC-3 on the expression of DKNIB gene.Western blotting was used to detect the expression of DKNIB protein and PI3K/AKT signaling pathway proteins.RESULTSCompared with chronic nasopharyngitis tissues,the expression of circRNA BCRC-3 in nasopharyngeal carcinoma tissues was significantly reduced(P<0.01).Compared with human immortalized nasopharyngeal epithelial cells,the expression of circRNA BCRC-3 in nasopharyngeal carcinoma cell lines was significantly reduced(P<0.01),and the expression in HONE-1cells was the lowest(P<0.01).Compared with the control group,the proliferation ability of HONE-1 cells in the experimental group decreased significantly from the second day(P<0.05),and the migration ability of HONE-1 cells was significantly reduced(P<0.01),and the expression of DKNIB in mRNA and protein levels of HONE-1 cells increased significantly(P<0.01),and the expression of PI3K/AKT signaling pathway proteins PIP3,PDK1,p-AKT,and AS160 significantly decreased.CONCLUSION circRNA BCRC-3 is low-expressed in nasopharyngeal carcinoma tissues and cell lines.Up-regulating circRNA BCRC-3 can inhibit the proliferation and migration of nasopharyngeal carcinoma HONE-1 cells.The mechanism may be that circRNA BCRC-3 up-regulates the expression of DKNIB gene and then inhibits PI3K/AKT signaling pathway transduction.