NE通过上调长链非编码RNA MALAT1抑制神经元增殖

NE inhibits neuronal proliferation by upregulating the expression of long noncoding RNA MALAT1

目的研究应激关键激素去甲肾上腺素(NE)对神经元细胞增殖的影响,分析长链非编码RNA肺癌转移相关转录本1(MALAT1)在其中的作用。方法通过不同浓度NE对HT22细胞进行干预,使用CCK-8检测各组细胞增殖活性,流式细胞术观察各组细胞周期分布,明确NE对细胞增殖的调控作用。采用实时荧光定量PCR(qRTPCR)法检测NE对神经元细胞中MALAT1表达的影响。利用慢病毒感染HT22细胞建立MALAT1稳定干扰细胞系,通过荧光表达水平和qRT-PCR验证干扰效果。将细胞分为shCtrl+DMSO、shCtrl+NE、shMALAT1+DMSO、shMALAT1+NE组,通过CCK-8实验和细胞周期实验检测干扰MALAT1的表达能否拮抗NE对HT22细胞的损伤作用。结果高浓度NE(100μmol/L)使S期细胞减少,G_(2)/M期细胞增加,从而抑制HT22细胞的增殖,同时qRTPCR结果显示,NE浓度为100μmol/L时可诱导神经元MALAT1表达显著升高;在HT22细胞中抑制MALAT1的表达可显著提高细胞的增殖能力,且干扰MALAT1的表达能拮抗NE对细胞的损伤作用。结论高浓度的NE通过上调MALAT1的表达水平抑制神经元细胞增殖。

Objective To study the effect of stress-associated hormone norepinephrine(NE)on neuronal cell proliferation and to analyze the role of long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1(MALAT1)in this process.Methods HT22 cells were intervened in by NE of different concentrations to study the regulatory effect of NE on cell proliferation.The proliferative activity and the cell cycle distribution of each group were detected using cell counting kit 8(CCK-8)and flow cytometry,respectively.The influence of NE on MALAT1 expressions in neuronal cells was detected by quantitative real-time fluorescence PCR(qRT-PCR).Cells that could stably intervene in MALAT1 and control cells were established through lentiviral infection,and the intervention was verified by fluorescence expression levels and qRT-PCR.The cells were divided into four groups:shCtrl+DMSO,shCtrl+NE,shMALAT1+DMSO,and shMALAT1+NE.The CCK-8 assay and cell cycle assay were performed to find out whether intervention in MALAT1 expression could antagonize the damage of NE to HT22 cells.Results A high concentration of NE(100μmol/L)decreased S-phase cells and increased G_(2)/M-phase cells,thus inhibiting the proliferation of HT22 cells.Meanwhile,qRT-PCR results showed that100μmol/L NE induced a significant increase of the expression of MALAT1 in HT22.Inhibition of MALAT1 expressions significantly increased the proliferation ability of HT22 cells.More importantly,the intervention with MALAT1 expressions could antagonize the damage of NE to HT22 cells.Conclusion High concentrations of NE can inhibit neuronal proliferation by upregulating the expression level of MALAT1.