MALAT1对Aβ1-42诱导PC12细胞损伤及PI3K/Akt通路的影响研究  

Study on the effects of MALAT1 on PC12 cell injury induced by Aβ1-42 and PI3K/Akt pathway

作　　者：王小言[1] 夏鹰[1] 彭俊[1] 金虎[1] 陈伟明[1] 陈晓东[1] 聂柳[1] 郑忠涛[1] WANG Xiaoyan;XIA Ying;PENG Jun;JIN Hu;CHEN Weiming;CHEN Xiaodong;NIE Liu;ZHENG Zhongtao(Department of Neurosurgery,Haikou Municipal Hospital,Haikou Hainan 570208,China)

机构地区：[1]海南省海口市人民医院神经外科,570208

出　　处：《中国神经免疫学和神经病学杂志》2020年第3期174-178,共5页Chinese Journal of Neuroimmunology and Neurology

基　　金：国家自然科学基金项目(NO.81360190);海南省卫生厅科研项目(NO.琼卫2011-57)。

摘　　要：目的探讨肺腺癌转移相关转录本1(MALAT1)对β-淀粉样蛋白1-42片段(Aβ1-42)诱导肾上腺嗜铬细胞瘤PC12细胞损伤及磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)通路的影响。方法体外培养PC12细胞,并随机分为正常对照组(正常生长PC细胞)、模型组(Aβ1-42诱导PC细胞损伤)、sh-MALAT1组(Aβ1-42组基础上转染MALAT1干扰序列)、sh-对照组(Aβ1-42组基础上转染MALAT1对照序列)。采用实时定量PCR(RT-qPCR)检测各组PC12细胞MALAT1表达水平,采用四甲基偶氮唑蓝(MTT)法检测PC12细胞增殖,采用流式细胞仪及AnnexinV-FITC/PI法体外检测PC12细胞凋亡,采用免疫印迹(WB)检测PC12细胞PI3K/Akt通路相关p-PI3K/PI3K、p-Akt/Akt蛋白及其下游凋亡相关蛋白Cleaved-caspase3/caspase3、B细胞淋巴瘤-2基因相关X蛋白(Bax)、B细胞淋巴瘤-2(Bcl-2)蛋白表达。结果与正常对照组比较,模型组、sh-对照组组PC12细胞存活率、p-PI3K/PI3K、p-Akt/Akt、Bcl-2蛋白水平低(均P<0.05),MALAT1表达水平、PC12细胞凋亡率、Bax、Cleaved-caspase3/caspase3蛋白表达水平高(均P<0.05);与模型组、sh-对照组比较,sh-MALAT1组MALAT1表达水平、细胞存活率及p-PI3K/PI3K、p-Akt/Akt、Bcl-2蛋白水平高(均P<0.05),Bax、Cleaved-caspase3/caspase3蛋白表达水平低(均P<0.05)。结论下调MALAT1表达可能通过激活PI3K/Akt通路,上调Bcl-2蛋白表达,下调Bax、Cleaved-caspase3/caspase3蛋白表达抑制Aβ1-42诱导的PC12损伤,发挥保护作用。Objective To study the effects of metastasis-associated lung adenocarcinoma transcript 1(MALAT1)on the injury of adrenal pheochromocytoma PC12 cells induced by amyloidβ-protein 1-42(Aβ1-42)fragment and the phosphatidylinositol 3 kinase/protein kinase B(PI3K/Akt)pathway.Methods PC12 cells were cultured in vitro and divided into four groups:a normal control group(normal PC cells),a model group(Aβ1-42 was used to induced PC cell damage),an sh-MALAT1 group(MALAT1 interference sequence was transfected on the basis of Aβ1-42 group),and an sh-control group(MALAT1 control sequence was transfected on the basis of Aβ1-42 group).Real time quantitative PCR(RT-qPCR)was used to detect the expression level of MALAT1 in PC12 cells.Methyl thiazolyl tetrazolium(MTT)was used to detect the proliferation of PC12 cells.Flow cytometry and AnnexinV-FITC/PI were used to detect the apoptosis of PC12 cells in vitro.Western blotting(WB)was used to detect the expressions of PI3K/Akt pathway related p-PI3K/PI3K,p-Akt/Akt proteins,and its downstream apoptosis related Cleaved-caspase3/caspase3,B-cell lymphoma-2 gene related X protein(Bax),B-cell lymphoma-2(Bcl-2)protein.Results Compared with the normal control group,PC12 cell survival rate,p-PI3K/PI3K,p-Akt/Akt and Bcl-2 protein levels in the model group and the sh-control group were lower(P<0.05),the expression levels of MALAT1,apoptosis rate of PC12 cells,Bax and Cleaved-caspase3/caspase3 proteins were all higher(P<0.05).Compared with the model group and the sh-Crtl group,MALAT1 expression level,cell survival rate,p-PI3K/PI3K,p-Akt/Akt and Bcl-2 protein levels in the sh-MALAT1 group increased(all P<0.05),and the expression levels of Bax and Cleaved-caspase3/caspase3 proteins decreased(P<0.05).Conclusions Down-regulating the expression of MALAT1 may up-regulate the expression of Bcl-2 protein and down-regulate the expressions of Bax and Cleaved caspase3/caspase3 proteins by activating the PI3K/Akt pathway,then inhibit the PC12 injury induced by Aβ1-42,and play the protective role.

关 键 词：肺腺癌转移相关转录本1 β-淀粉样蛋白 PC12细胞 1-磷脂酰肌醇3-激酶 蛋白激酶B 信号通路 阿尔茨海默病 

分 类 号：R742.89[医药卫生—神经病学与精神病学] Q257[医药卫生—临床医学]