长链非编码RNAMALAT1调控星形胶质细胞增殖和凋亡及对MAPK/ERK1,2信号通路的影响  

Long non⁃coding RNA MALAT1 regulates astrocyte proliferation and apoptosis and affects MAPK/ERK1,2 signaling pathway

作　　者：胡辉 王雪[1] 吴雨涵 董华丰 张玲 韦爱君 谢方 赵云[1] 孙兆炜 钱令嘉 HU Hui;WANG Xue;WU Yuhan;DONG Huafeng;ZHANG Ling;WEI Aijun;XIE Fang;ZHAO yun;SUN Zhaowei;QIAN Lingjia(Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China)

机构地区：[1]军事科学院军事医学研究院,北京100850

出　　处：《军事医学》2024年第5期347-354,共8页Military Medical Sciences

基　　金：国家自然科学基金项目(82271553);北京市自然科学基金项目(5222033)。

摘　　要：目的探讨肺腺癌转移相关转录本1(MALAT1)的表达对星形胶质细胞(C8-D1A)增殖和凋亡,以及丝裂原活化蛋白激酶(MAPK)/细胞外信号调节激酶(ERK1,2)通路的影响。方法通过慢病毒载体和质粒载体在C8-D1A细胞中分别敲低及过表达MALAT1基因,用实时荧光定量PCR(qPCR)法检测敲低及过表达效率、细胞增殖相关标志物Ki67、微染色体维持蛋白2(MCM2)、细胞核增殖抗原(PCNA)、凋亡相关蛋白含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)、B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)的表达变化,CCK-8、流式细胞术检测敲低及过表达MALAT1后对C8-D1A细胞增殖和凋亡的影响。免疫荧光染色法检测Caspase-3和Ki67蛋白表达,Western印迹法检测细胞凋亡相关蛋白Caspase-3、Bax、Bcl-2及ERK1/2、p-ERK1/2、p38MAPK、p-p38MAPK蛋白表达变化情况。结果与对照组相比,过表达MALAT1抑制细胞增殖并诱导细胞凋亡,其G0/G1期、G2/M期比例升高,S期比例降低,Caspase-3蛋白表达升高,p-ERK1/2蛋白表达降低,Ki67、PCNA mRNA表达量降低;而敲低MALAT1后细胞增殖显著增加并抑制细胞凋亡,其S期比例增高、G2/M期比例降低,Caspase-3、Bax蛋白表达降低,p-ERK1/2、p-p38MAPK蛋白表达升高,Ki67、MCM2、PCNA mRNA表达量增加(P<0.05)。结论MALAT1基因可能通过调控MAPK/ERK1,2信号通路进而促进星形胶质细胞凋亡并抑制其增殖。Objective To investigate the effect of MALAT1 expressions on cell proliferation and apoptosis in astrocytes by regulating mitogen-activated protein kinase(MAPK)/extracellular signal-regulated kinase(ERK1,2)pathway.Methods The MALAT1 gene was knocked down and over-expressed in C8-D1A cells by lentiviral and plasmid vectors,respectively.The expressions of MALAT1,cell proliferation-related markers(Ki67,MCM2,PCNA)and apoptosis-related proteins(Caspase-3,Bax,Bcl-2)weredetectedbyquantitativereal-timepolymerasechainreaction(qPCR).CCK-8assayand flow cytometry were used for cell proliferation and apoptosis in C8-D1A cells.Immunofluorescence was adopted to detect the protein expressions of Caspase-3 and Ki67.Western blotting was used to detect the protein expressions of Caspase-3,Bax,Bcl-2,ERK1/2,p-ERK1/2,p38MAPK and p-p38MAPK.Results Compared with the control group,over-expressed MALAT1 inhibited cell proliferation and induced cell apoptosis in C8-D1A cells while the knockdown of MALAT1 significantly enhanced cell proliferation and anti-apoptotic ability in C8-D1A cells.The proportion of C8-D1A cells in G0/G1-phase and G2/M-phase was higher than in the control group as evidenced by flow cytometry,but was lower in S-phase.Meanwhile,datashowedthatCaspase-3wasincreasedwhilep-ERK1/2wasdecreasedintermsofprotein levels.The mRNA expressions of Ki67 and PCNA were decreased.After knockdown of MALAT1,the proportion of C8-D1A cells in S-phase was higher,but was lower in G2/M-phase.The protein expressions of Caspase-3 and Bax decreased while those of p-ERK1/2 and p-p38MAPK increased.The mRNA expressions of Ki67,MCM2 and PCNA were increased.The differences were all statistically significant(P<0.05).Conclusion MALAT1 promotes astrocyte apoptosis and inhibits proliferation by regulating the MAPK/ERK1,2 signaling pathway.

关 键 词：肺腺癌转移相关转录本1 星形胶质细胞 丝裂原活化蛋白激酶 细胞外信号调节激酶 细胞凋亡 细胞增殖 

分 类 号：R741.02[医药卫生—神经病学与精神病学]