磷脂酶CB1对胶质瘤U87细胞增殖和侵袭的影响  

EFFECT OF PHOSPHOLIPASE CB1 ON THE PROLIFERATION AND INVASION OF GLIOMA U87 CELLS

作　　者：赵鹏[1] 孙树凯[1] 翟玉娥[1] 田清武[1] 周廷廷 李靖[2] ZHAO Peng;SUN Shukai;ZHAI Yu′e;TIAN Qingwu;ZHOU Tingting;LI Jing(Department of Clinical Laboratory, The Affiliated Hospital of Qingdao University, Qingdao 266003, China)

机构地区：[1]青岛大学附属医院检验科,山东青岛266003 [2]青岛大学附属医院肾内科,山东青岛266003

出　　处：《精准医学杂志》2020年第6期528-532,共5页Journal of Precision Medicine

摘　　要：目的探讨磷脂酶CB1(PLCB1)对胶质瘤U87细胞增殖和侵袭的影响。方法选取50例新鲜神经胶质瘤组织(胶质瘤组)及其癌旁非肿瘤组织(癌旁组),每份组织样本分为两份,一份采用免疫组化法分析检测组织中PLCB1的表达,一份采用实时荧光定量PCR(RT-qPCR)和Western-blot实验检测组织中PLCB1的表达。体外培养胶质瘤U87细胞,将PLCB1 siRNA(PLCB1 siRNA组)和Con siRNA(Con siRNA组)分别转染至神经胶质瘤U87细胞后,采用CCK-8分析检测PLCB1对两组细胞增殖的影响;采用Transwell实验和划痕实验检测PLCB1对两组细胞侵袭和迁移的影响;采用Western-blot实验检测PLCB1 siRNA组和Con siRNA组中细胞外调节蛋白激酶(ERK)以及磷酸化细胞外调节蛋白激酶1/2(p-ERK1/2)的表达水平。结果qPCR检测结果显示,胶质瘤组中PLCB1 mRNA的表达水平明显高于癌旁组(t=131.29,P<0.01);免疫组化和Western-blot检测结果显示,胶质瘤组中PLCB1蛋白表达量明显高于癌旁组(t=100.53、50.25,P<0.01)。CCK-8分析结果显示,与Con siRNA组相比,PLCB1 siRNA组中神经胶质瘤U87细胞的增殖能力明显抑制(F=61.16,P<0.05)。Transwell和划痕实验结果显示,与Con siRNA组相比,PLCB1 siRNA组细胞侵袭和迁移能力明显受到抑制(F=29.96、55.71,P<0.05)。Western-blot实验结果显示,PLCB1 siRNA可以抑制p-ERK1/2的表达(F=17.98、24.67,P<0.01)。结论胶质瘤组织中PLCB1蛋白高表达,PLCB1通过激活ERK通路促进胶质瘤细胞的增殖、侵袭和迁移。Objective\To investigate the effect of phospholipase CB1(PLCB1)on the proliferation and invasion of glioma U87 cells.\Methods\A total of 50 fresh glioma tissue samples(glioma tissue group)and their adjacent non-tumor tissue samples(adjacent tissue group)were selected,and each tissue sample was divided into two parts,of which one part was used for immunohistochemistry to measure the expression of PLCB1 and the other part was used for quantitative real-time PCR(RT-qPCR)and Western blot to measure the expression of PLCB1.Glioma U87 cells were cultured in vitro.After PLCB1 siRNA(PLCB1 siRNA group)and Con siRNA(Con siRNA group)were transfected into glioma U87 cells,CCK-8 assay was used to observe the effect of PLCB1 on the proliferation of U87 cells;Transwell assay and wound healing assay were used to evaluate the effect of PLCB1 on cell invasion and migration;Western blot was used to measure the expression of extracellular signal-regulated kinase(ERK)and phosphorylated extracellular signal-regulated kinase 1/2(p-ERK1/2).\Results\The RT-qPCR results showed that the glioma tissue group had significantly higher mRNA expression of PLCB1 than the adjacent tissue group(t=131.29,P<0.01),and the results of immunohistochemistry and Western blot showed that the glioma tissue group had significantly higher protein expression of PLCB1 than the adjacent tissue group(t=100.53,50.25,P<0.01).The results of CCK-8 assay showed that compared with the Con siRNA group,the PLCB1 siRNA group had significant inhibition of the proliferation of glioma U87 cells(F=61.16,P<0.05).Transwell assay and wound healing assay showed that compared with the Con siRNA group,the PLCB1 siRNA group had significant inhibition of the invasion and migration abilities of glioma cells(F=29.96,55.71,P<0.05).Western blot showed that PLCB1 siRNA inhibited the expression of p-ERK1/2(F=17.98,24.67,P<0.01).\Conclusion\PLCB1 protein is highly expressed in glioma tissue,and PLCB1 promotes the proliferation,invasion,and migration of glioma cells by activating the ERK path

关 键 词：磷脂酶CB 神经胶质瘤 细胞增殖 细胞运动 肿瘤侵润 细胞外调节蛋白激酶 

分 类 号：R730.264[医药卫生—肿瘤]