CASC11通过调控miR-641的表达对脑胶质瘤细胞恶性生物学行为的作用机制研究

The mechanism of CASC11 on the malignant biological behavior of glioma cells by regulating the expression of miR-641

目的 探讨长链非编码RNA癌症易感候选基因11(Long non-coding RNA cancer susceptibility candidate gene 11,lncRNA CASC11)通过调控微小核糖核酸(MicroRNA,miR)-641的表达对脑胶质瘤细胞恶性生物学行为的作用机制。方法 逆转录-聚合酶链反应(Reverse transcription-polymerase chain reaction,RT-PCR)检测脑胶质瘤细胞株U251,U87,MGR3,UWR7和A172中CASC11表达水平及小干扰RAN CASC11(Small interfering RNA CASC11,si-CASC11)转染U87细胞后CASC11,miR-641表达水平;在人脑胶质瘤细胞U87中敲除CASC11后用四甲基偶氮唑盐(Methyl thiazolyl tetrazolium,MTT)法、侵袭小室实验(Transwell assay)和细胞划痕实验检测U87细胞增殖、侵袭和迁移能力;双荧光素酶报告基因验证CASC11和miR-641之间的靶向关系。结果 在U87细胞中CASC11表达水平最高,故选U87细胞进行后续实验;敲除CASC11后可抑制U87细胞增殖、侵袭和迁移能力,降低U87细胞中的CASC11 mRNA相对表达水平,升高miR-641 mRNA相对表达水平;CASC11和miR-641之间存在靶向关系。结论 敲除CASC11后可靶向负调控miR-641表达,抑制脑胶质瘤细胞增殖、侵袭和迁移的恶性生物学行为,可能是脑胶质瘤的分子靶点。

Objective To explore the mechanism of long non-coding RNA cancer susceptibility candidate gene 11(lncRNA CASC11) in regulating the expression of microribonucleic acid(miR)-641 on the malignant biological behavior of glioma cells.Methods The technique of reverse Transcription-Polymerase Chain Reaction(RT-PCR) is used to detect the transcriptional level of CASC11 in glioma cell lines U251,U87,MGR3,UWR7 and A172,and the levels of CASC11 and miR-641 were detected aftersi-CASC11 is transfected into U87 cells.CASC11 gene was also knocked out in human glioma cell U87,and then MTT method,transwell chamber and cell scratch test were used to detect U87 cell proliferation,invasion and migration ability.The targeting relationship between CASC11 and miR-641 was determined by using dual luciferase reporter gene.Results CASC11 has the highest expression level in U87 cells,and so U87 cells were selected for subsequent experiments.Knocking out CASC11 gene could inhibit the proliferation,invasion and migration ability of U87 cells,reduce the relative level of CASC11 mRNA in U87 cells,and increase the relative level of miR-641 mRNA.There was a targeting relationship between CASC11 and miR-641.Conclusion Knocking out of CASC11 gene can target and negatively regulate the expression of miR-641,inhibit the malignant biological behavior of glioma cell proliferation,invasion and migration,and CASC11 may be a novel molecular target of glioma therapy.