miR-138靶向作用Sema4C对脑胶质瘤细胞侵袭转移的影响

Effects of miR-138 on invasion of brain glioma cells through targeting Sema4C

目的研究微小RNA-138（miR-138）对脑胶质瘤细胞侵袭转移的影响及其机制。方法实时荧光定量聚合酶链反应（qRT-PCR）检测miR-138在60例脑胶质瘤组织及其癌旁组织中的表达，分析miR-138表达与脑胶质瘤分级的关系；体外对脑胶质瘤细胞株U251和U87分别转染miR-138类似物（miR-138 mimic）及阴性对照（miR-NC）；qRT-PCR检测miR-138表达情况；Transwell实验检测U251和U87细胞侵袭转移能力；Western blotting检测miR-138靶向蛋白轴突导向蛋白4C（Sema4C）的表达。结果脑胶质瘤组织中miR-138表达水平（2.46±1.07）较癌旁正常脑组织（4.83±1.16）明显降低（t=-11.631，P＜0.001），且miR-138表达与肿瘤恶性程度呈负相关（r=-0.563，P=0.001）。转染miR-138 mimic组细胞miR-138表达（U251：3.96±0.16；U87：4.43±0.96）明显高于转染miR-NC组（U251：2.32±0.36；U87：2.58±0.62）（t=7.253，P＜0.001；t=8.872，P＜0.001）。miR-138 mimic组（U251：89±9；U87：95±10）细胞侵袭转移能力明显低于转染miR-NC组（U251：206±15；U87：240±20）（t=36.629，P＜0.001；t=35.521，P＜0.001）。miR-138 mimic组细胞Sema4C表达（U251：0.41±0.06；U87：0.36±0.03）明显低于miR-NC组（U251：1.01±0.08；U87：1.03±0.13）（t=-32.862，P＜0.001；t=-27.512，P＜0.001）。结论上调miR-138表达可抑制脑胶质瘤细胞侵袭转移，可能与负向调节Sema4C表达有关。

ObjectiveTo explore the effects and mechanism of microRNA-138 （miR-138） on brain glioma cells invasion and migration. MethodsThe expression of miR-138 was detected by real time quantitative polymerase chain reaction （qRT-PCR） in 60 cases of glioma tissues and paracarcinoma tissues, and the relationship between miR-138 expression and glioma grading was analyzed. Human glioma cells line U87 and U251 were transfected with miR-138 mimic and negative control （miR-NC）. The expression of miR-138 was detected by qRT-PCR. The migration and invasion abilities were tested by Transwell assay, and the expression of semaphoring 4C （Sema4C） protein was tested by Western blotting. ResultsThe expression level of miR-138 in glioma tissues （2.46±1.07） was significantly lower than that in normal brain tissues （4.83±1.16, t=-11.631, P〈0.001）, and miR-138 expression was negatively correlated with tumor grade （r=-0.563, P=0.001）. The expression level of miR-138 in cells was significantly higher after being transfected with miR-138 mimic （U251： 3.96±0.16; U87： 4.43±0.96） than miR-NC （U251： 2.32±0.36; U87： 2.58±0.62, t=7.253, P〈0.001; t=8.872, P〈0.001）. The ability of invasion and migration were lower after being transfected with miR-138 mimic （U251： 89±9; U87： 95±10） than miR-NC （U251： 206±15; U87： 240±20, t=36.629, P〈0.001; t=35.521, P〈0.001）. The expression of Sema4C protein was lower after being transfected with miR-138 mimic （U251： 0.41±0.06; U87： 0.36±0.03） than miR-NC （U251： 1.01±0.08; U87： 1.03±0.13, t=-32.862, P〈0.001; t=-27.512, P〈0.001）. ConclusionThe upregulated expression of miR-138 can suppress glioma cells migration and invasion, which might be related to the negative regulation expression of Sema4C protein.