miR-497对恶性脑膜瘤细胞增殖的影响

Effect of miR-497 on proliferation of malignant meningioma cells

目的探讨miR-497对恶性脑膜瘤细胞增殖的影响及作用机制。方法收集2019年1~9月手术切除的女性脑膜瘤组织标本10例,其中恶性脑膜瘤5例(WHO分级Ⅲ级),良性脑膜瘤5例(WHO分级Ⅰ级),采用RNA-seq法和qRT-PCR检测脂肪酸合成酶(FASN)mRNA和miR-497表达水平。体外培养恶性脑膜瘤细胞株IOMM-Lee细胞,设置空白组(不做任何处理)、miR-NC组(转染negative control质粒)、miR-497-m组(转染miR-497 mimics)和miR-497-i组(转染miR-497 inhibitor),MTT法检测细胞增殖能力,检测细胞上清液乳酸水平和细胞ATP含量,免疫印迹法检测细胞FASN蛋白表达水平。结果恶性脑膜瘤组织FASN mRNA水平明显高于良性脑膜瘤组织(P<0.05),而miR-497水平则明显低于良性脑膜瘤组织(P<0.05)。与空白组和miR-NC组相比,miR-497-m组细胞增殖能力、细胞上清液乳酸水平、细胞ATP含量以及FASN蛋白表达水平均明显降低(P<0.05);而miR-497-i组细胞增殖能力、细胞上清液乳酸水平、细胞ATP含量以及FASN蛋白表达水平均明显升高(P<0.05)。结论恶性脑膜瘤组织miR-497呈低表达,通过靶向促进FASN表达,提高肿瘤细胞内脂肪酸代谢水平,促进肿瘤细胞增殖。

Objective To investigate the effect of miR-497 on the proliferation of malignant meningioma cells and its mechanism.Methods The levels of fatty acid synthase(FASN)mRNA and miR-497 were detected by RNA-seq and qRT-PCR in meningioma tissues obtained from 10 female patients with meningioma including 5 malignant and 5 benign meningiomas.IOMM-Lee cells were cultured in vitro and divided into blank group,miR-NC group(transfected with negative control),miR-497-m group(transfected with miR-497 mimics)and miR-497-i group(transfected with miR-497 inhibitor).The cell proliferation ability was detected by MTT method;lactic acid in cell supernatant and cell ATP contents were detected;FASN protein expression were measured using western blotting.Results The mRNA level of FASN was signifcantly higher and the miR-497 level was signifcantly lower in malignant meningioma tissues than benign meningioma tissues(P<0.05).Compared with blank and miR-NC groups,the cell proliferation activity,lactate acid level in the cell supernatant,cell ATP content and FASN protein expression level significantly decreased in miR-497-m group(P<0.05),while significantly increased in miR-497-i group(P<0.05).Conclusion The expression level of miR-497 in malignant meningioma tissue is low,which may be by promoting FASN expression and increasing the fatty acid metabolism level in tumor cells to promote tumor cell proliferation.