LncRNA MYU对胶质瘤细胞周期分布、增殖、转移和凋亡以及PI3K/Akt信号通路的影响

摘要 目的：探讨长链非编码RNA（LncRNA）MYU对胶质瘤细胞周期分布、细胞增殖、迁移、侵袭和凋亡的影响，并初步探讨其作用机制。方法：实时荧光定量PCR（RT-qPCR）检测人脑正常胶质细胞HEB和胶质瘤细胞（U-251MG、A172、SHG139）中LncRNA MYU的表达情况。选取SHG139细胞，分为正常对照（NC）组、si-con组、si-LncRNA MYU组进行转染实验，行RT-qPCR检测转染效果。分别采用流式细胞术、细胞计数试剂盒（CCK-8）、Transwell实验检测沉默LncRNA MYU对SHG139细胞周期分布和凋亡、细胞增殖、细胞迁移和侵袭的影响。蛋白免疫印迹（Western blot）法检测基质金属蛋白酶2（MMP-2）、MMP-9、裂解的半胱氨酸天冬氨酸蛋白酶3（Cleaved caspase-3）、Cleaved caspase-9以及磷脂酰肌醇-3-羟激酶/蛋白激酶B（PI3K/Akt）信号通路相关蛋白表达情况。结果：LncRNA MYU在胶质瘤细胞株中比人脑正常胶质细胞中的表达水平显著升高（P＜0.05），因此选择表达量最高的SHG139细胞进行转染实验。沉默LncRNA MYU能够显著诱导SHG139细胞G0-G1期阻滞、抑制细胞增殖、迁移和侵袭并诱导细胞凋亡（P＜0.05）。沉默LncRNA MYU可显著抑制MMP-2、MMP-9、p-PI3K和p-AKT表达并促进Cleaved caspase-3、Cleaved caspase-9表达（P＜0.05）。结论：沉默LncRNA MYU可诱导胶质瘤细胞G0-G1期阻滞，抑制细胞增殖、迁移和侵袭，促进细胞凋亡，其机制可能与抑制PI3K/AKT信号通路有关。

Effects of LncRNA MYU on the Cycle Distribution, Proliferation, Metastasis and Apoptosis of Glioma Cells and the PI3K/Akt Signaling Pathway

ABSTRACT Objective: To investigate the effects of long-chain non-coding RNA (LncRNA) MYU on the cycle distribution, cell proliferation, migration, invasion, and apoptosis of glioma cells, and to preliminarily explore its mechanism of action. Methods: The expression of LncRNA MYU in human brain normal glial cells HEB and glioma cells (U-251MG, A172, SHG139) was detected by real- time fluorescent quantitative PCR (RT-qPCR). SHG139 cells were selected and divided into normal control (NC) group, si-con group, and si-LncRNA MYU group for transfection experiments, and RT-qPCR was applied to detect the transfection effect. The effects of silencing LncRNA MYU on the cell cycle distribution and apoptosis, cell proliferation and, cell migration and invasion of SHG139 cells were detected respectively by flow cytometry, cell counting kit (CCK-8), and Transwell experiments. Western blot was used to detect the expression of matrix metalloproteinase (MMP-2), MMP-9, Cleaved caspase-3, Cleaved caspase-9, and Phosphatidylinositol-3-hydroxykinase/protein kinase B (PI3K/Akt) signaling pathway-related proteins. Results: The expression of LncRNA MYU in glioma cell lines was significantly higher than that in normal human brain glial cells(P<0.05), thereby SHG139 cells with the highest expression levels were selected for transfection experiments. Silencing LncRNA MYU can significantly induce G0-G1 phase arrest, inhibit the proliferation, migration and invasion, and induce apoptosis in SHG139 cells(P<0.05). Silencing LncRNA MYU can significantly inhibit the expression of MMP-2, MMP-9, p-PI3K and p-AKT, and promote the expression of Cleaved caspase-3 and Cleaved caspase-9(P<0.05). Conclusion: Silencing LncRNA MYU can induce G0-G1 phase arrest, inhibit cell proliferation, migration and invasion, and promote cell apoptosis in glioma cells, and its mechanism may be related to the inhibition of PI3K/AKT signaling pathway.