摘要目的:观察微小RNA(miRNA，miR)-4295靶向调控多亮氨酸重复区免疫球蛋白样蛋白1(LRIG1)对脑胶质瘤细胞增殖、侵袭和上皮-间充质转化(EMT)的影响。方法:选取2018年6月到2020年4月经手术切除的120例脑胶质瘤及癌旁组织作为研究对象，采用荧光定量聚合酶链反应(PCR)分析胶质瘤组织和癌旁组织中miR-4295表达水平。采用慢病毒在脑胶质瘤细胞系U251建立miR-4295沉默细胞系(实验组)和对照细胞系(对照组)。采用细胞计数试剂盒(CCK-8)分析两组细胞增殖能力；采用Transwell分析细胞侵袭能力；采用蛋白质印迹法(Western blot)分析对照组和实验组EMT标志物表达变化。生物信息学和双荧光素酶报告基因分析miR-4295基因，并分析对照组和实验组细胞靶蛋白表达水平。组间数据比较采用 t检验。 结果:与癌旁组织(1.29±0.20)比较，脑胶质瘤组织中miR-4295表达水平(2.81±0.19)显著增加，差异有统计学意义( t＝2.809， P<0.05)。与对照组(1.07±0.15)比较，实验组细胞miR-4295表达水平(0.38±0.11)显著下调，差异有统计学意义( t＝2.441， P<0.05)。与对照组细胞48 h吸光度( A)值(1.61±0.25)比较，实验组细胞48 h A值(1.22±0.18)显著下降，差异有统计学意义( t＝2.091， P<0.05)。与对照组细胞[(89.27±9.10)个]比较，实验组侵袭细胞数量[(32.49±5.94)个]显著下降，差异有统计学意义( t＝4.109， P<0.05)。与对照组细胞N-钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)蛋白表达水平(1.13±0.13、1.20±0.19)比较，实验组细胞N-cadherin和Vimentin蛋白表达水平(0.30±0.11、0.37±0.12)显著下调，差异有统计学意义( t＝3.091、2.861， P<0.05)。生物信息学和双荧光素酶报告基因显示LRIG1是miR-4295靶基因。癌旁组织中LRIG1相对表达水平(1.05±0.22)比较，肿瘤组织LRIG1相对表达水平(0.23±0.11)显著下调，差异有统计学意义( t＝3.021， P<0.05)。与对照组细胞LRIG1相对表达水平(0.35±0.13)比较，实验组细胞LRIG1相对表达水平(0.89±0.17)显著增加，差异有统计学意义( t＝2.198， P<0.05)。 结论:miR-4295通过靶向LRIG1调控着脑胶质瘤细胞增殖、侵袭和EMT。

abstractsObjective:To investigate the effects of miR-4295 on proliferation, invasion and epithelial mesenchymal transition of glioma cells by targeting leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1).Methods:120 cases gliomas and adjacent tissues from June 2018 to April 2020 were selected as the research objects. The expression level of miR-4295 in gliomas and adjacent tissues was analyzed by fluorescence quantitative polymerase chain reaction (PCR). MiR-4295 silencing cell line (experimental group) and control cell line (control group) were established by lentivirus in U251 glioma cell line. The proliferation ability of the two groups was analyzed by cell counting kit-8 (CCK-8). The invasion ability was analyzed by Transwell. The expression of epithelial mesenchymal markers in the control group and the experimental group was analyzed by Western blotting. The miR-4295 gene was analyzed by bioinformatics and double luciferase reporter gene, and the expression level of cell target protein in control group and experimental group was analyzed by Western blotting. T test was used to compare the data between groups, and the difference was statistically significant ( P<0.05). Results:The expression of miR-4295 in gliomas (1.29±0.20) was significantly higher than that in adjacent tissues (2.81±0.19, t=2.809, P<0.05). Compared with the control group (1.07±0.15), the expression level of miR-4295 in the experimental group (0.38±0.11) significantly reduced ( t=2.441, P<0.05). Compared with the control group (1.61±0.25), the 48 h absorbance value (1.22±0.18) of the experimental group significantly reduced ( t=2.091, P<0.05). Compared with the control group [ (89.27±9.10) cells], the number of invasive cells in the experimental group [ (32.49±5.94) cells] significantly decreased ( t=4.109, P<0.05). The expression levels of N-cadherin and Vimentin in the experimental group (0.30±0.11, 0.37±0.12) were significantly lower than those in the control group (1.13±0.13, 1.20±0.19, t=3.091, 2.861, P<0.05). Bioinformatics and double luciferase reporter gene showed that LRIG1 was the target gene of miR-4295. The relative expression level of LRIG1 in the adjacent tissues (1.05±0.22) was significantly lower than of tumor tissues (0.23±0.11, t=3.021, P<0.05). Compared with the control group (0.35±0.13), the relative expression level of LRIG1 in the experimental group significantly increased (0.89±0.17, t=2.198, P<0.05). Conclusion:MiR-4295 regulates the proliferation, invasion and epithelial mesenchymal transition of glioma cells by targeting LRIG1.