LASP1与MTDH互相调控介导121例人脑胶质瘤组织及细胞株异常增殖探讨  

Molecular mechanism of LASP1 and MTDH regulation and regulation of abnormal proliferation of human glioma

作　　者：罗穆云[1] 肖秋香[2] 廖伟[1] 易仁辉 李伟松[2] 刘四君[2] LUO Mu-yun;XIAO Qiu-xiang;LIAO Wei;YI Ren-hui;LI Wei-song;LIU Si-jun(First Affiliated Hospital of Gannan Medical College,Ganzhou 341099,China)

机构地区：[1]赣南医学院第一附属医院神经外科,江西赣州341099 [2]赣南医学院第一附属医院病理科,江西赣州341099

出　　处：《中华肿瘤防治杂志》2021年第1期31-41,共11页Chinese Journal of Cancer Prevention and Treatment

基　　金：江西省教育厅科学技术一般项目(GJJ180806);江西省教育厅科技课题(160993)。

摘　　要：目的探讨LASP1和MTDH互相调控介导人脑胶质瘤异常增殖的分子机制。方法收集赣南医学院第一附属医院2013-05-19-2018-03-21首诊并进行手术治疗的121例脑胶质瘤患者瘤组织及其癌旁正常脑组织,使用定量聚合酶链式反应(qPCR)、蛋白质印迹法和免疫组化法检测LASP1和MTDH表达。构建4个实验组,采用U251细胞株进行转染。第1组转染pcDNA-LASP1与pshRNA-MTDH,第2组转染pshRNA-LASP1与pcDNA-MTDH,第3组转染pshRNA-LASP1与pshRNA-MTDH,第4组转染无意义序列。采用qPCR和蛋白质印迹法测量4组LASP1、MTDH、miR-203a-3p、Brk1、基质金属蛋白酶-2(MMP-2)和MMP-9的表达,使用甲基化PCR测量甲基化MTDH的表达,通过克隆形成、划痕愈合和Transwell试验检测4组细胞增殖、迁移和侵袭能力。对U251细胞株使用不同浓度PDTC与PMA培养后,通过qPCR和蛋白质印迹法检测LASP1和MTDH的表达。2组间均值比较用t检验,多组均值间比较用单因素方差分析,两两多重比较用LSD检验;2组间率的比较则采用χ^(2)检验;双定量变量相关用Pearson分析。结果人体组织标本测定指标LASP1和MTDH在121例胶质瘤组织中表达量分别为6.31±1.70和5.89±1.24,在正常组织中表达量分别为4.27±1.07和3.40±0.99(t=5.224,P=0.032;t=3.561,P=0.017),两者表达量与胶质瘤恶性程度正相关(r=0.535,P=0.036;r=0.610,P=0.042)。LASP1与MTDH之间呈正相关(r=0.692,P=0.041),LASP1和MTDH中任意一方表达加强时均会导致另一方表达也同步增加,反之亦然。体外细胞实验结果显示,LASP1和MTDH通过增加Brk1、MMP-2和MMP-9的表达,从而增加胶质瘤细胞增殖、迁移和侵袭能力。MTDH影响程度高于LASP1(t=4.526,P=0.035),LASP1通过调节miR-203a-3p表达从而对MTDH翻译过程进行调控(t=6.391,P=0.023),而MTDH则通过核转录因子κB信号通路对LASP1转录过程进行调节。结论LASP1通过miR-203a-3p对MTDH翻译过程进行调控,而MTDH通过NF-κB信号通路对LASP1的转录过程进�Objective To investigate the molecular mechanism that LASP1 and MTDH interact with each other to mediate human glioma abnormal proliferation.Methods Totally 121 cases of glioma patients and normal brain tissues adjacent to the cancer were collected from the First Affiliated Hospital of Gannan Medical College of Jiangxi Province from May 19,2013 to March 21,2018,and qPCR,Western blot,and immunohistochemical methods were used to detect LASP1 MTDH expression.Four experimental groups were constracted:transfection with U251 cell line,group 1 transfected with pcDNA-LASP1 and pshRNA-MTDH,group 2 transfected with pshRNA-LASP1 and pcDNA-MTDH,and group 3 transfected with pshRNA-LASP1 and pshRNA-MTDH,group 4 transfected with nonsense sequences.The qPCR and Western blot methods were used to measure the expression of 4 groups of LASP1,MTDH,miR-203 a-3 p,Brk1,matrix metalloproteinase-2(MMP-2)and MMP-9.The methylated PCR was used to measure the expression of methylated MTDH,and clone formation and scratch healing were performed.The Transwell test was used to detect the cell proliferation,migration and invasion ability of the four groups.After U251 cell lines were cultured with different concentrations of PDTC and PMA,the expressions of LASP1 and MTDH were detected by qPCR and Western blot.The mean values of two groups were compared using the t-test,the mean values of multiple groups were compared using the one-way analysis of variance,and the mean values of two groups were compared using the LSD test.Theχ^(2) test was used to compare the rates between the two groups.The correlation between the two disordered variables was analyzed by Pearson.Results The expression levels of LASP1 and MTDH in glioma tissues(6.31±1.70,5.89±1.24)were much higher than those in normal tissues(4.27±1.07,3.40±0.99;t=5.224,P=0.032;t=3.561,P=0.017)and the expression levels of the two and the gel.There was a positive correlation between the degree of malignant tumors(r=0.535,P=0.036;r=0.610,P=0.042).By correlation analysis,there was a positive corre

关 键 词：LASP1 MTDH 胶质瘤 miR-203a-3p 核转录因子ΚB 

分 类 号：R739.41[医药卫生—肿瘤]