miR-30d-5p通过DLL4/NOTCH信号通路抑制胶质瘤细胞增殖及血管生成

miR-30d-5p inhibits glioma cell proliferation and angiogenesis through DLL4/NOTCH signaling pathway

目的探索miR-30d-5p对胶质瘤细胞的增殖、血管生成的影响以及可能机制。方法采集胶质瘤患者临床样本共20例,荧光定量PCR法检测胶质瘤患者的癌组织及正常脑组织中miR-30d-5p的表达以及不同胶质瘤细胞中miR-30d-5p的表达。荧光素酶报告实验检测miR-30d-5p与DLL4的靶向性。将裸鼠随机分为对照组(NC组)与miR-30d-5p mimics组,荧光定量PCR法及Western blot法检测miR-30d-5pNC、miR-30d-5p mimics转染后胶质瘤细胞DLL4的表达;裸鼠成瘤实验观察miR-30d-5p mimics对成瘤的影响;免疫荧光检测瘤体CD31表达,Western blot法检测瘤体中DLL4、NOTCH1、NOTCH2的表达。结果miR-30d-5p在胶质瘤患者及胶质瘤细胞中表达降低;DLL4是miR-30d-5p的一个作用靶点,miR-30d-5p可抑制DLL4的表达并促进CD31的表达;miR-30d-5抑制瘤体生长并抑制瘤体中DLL4、NOTCH1、NOTCH2的表达。结论miR-30d-5p可通过作用DLL4/NOTCH信号通路来抑制胶质瘤细胞增殖及血管生成,抑制肿瘤生长。

Objective To explore the effects and possible mechanisms of miR-30d-5p on the proliferation and angiogenesis of glioma cells.Methods A total of 20 clinical samples from glioma patients were collected.The expression of miR-30d-5p in cancer tissues and normal brain tissues of glioma patients and the expression of miR-30d-5p in different glioma cells were detected by fluorescence quantitative PCR.The targeting of miR-30d-5p and DLL4 was detected by luciferase reporting assay.Nude mice were randomly divided into control group(NC group)and miR-30d-5pmimics group.The expression of DLL4 in glioma cells transfected by miR-30d-5pNC and miR-30d-5pmimics was detected by fluorescence quantitative PCR and Western blot.The effect of miR-30d-5p mimics on tumor formation was observed in nude mice.The expression of CD31 in the tumor was detected by immunofluorescence,and the expressions of DLL4,NOTCH1and NOTCH2 in the tumor were detected by Western blot.Results The expression of miR-30d-5p was decreased in glioma patients and glioma cells.DLL4 is a target of miR-30d-5p,which can inhibit the expression of DLL4 and promote the expression of CD31.miR-30d-5 inhibited the tumor growth and the expression of DLL4,NOTCH1 and NOTCH2 in the tumor.Conclusion miR-30d-5p can inhibit the proliferation and angiogenesis of glioma cells and inhibit tumor growth by acting on the DLL4/NOTCH signaling pathway.