补骨脂素通过miR-101/PI3K/Akt轴对骨肉瘤细胞侵袭、转移的影响  

Effect of psoralen on invasion and metastasis of osteosarcoma cells through miR101/PI3K/Akt axis

作　　者：史博 杨健 张立喜[3] SHI Bo;YANG Jian;ZHANG Lixi(Department of Orthopedics,Cangzhou People’s Hospital,Cangzhou 061000,China;Department of Orthopedics,Cangzhou Central Hospital,Cangzhou 061001;Department of Orthopedics,Fengnan District Hospital of Tangshan City,Tangshan 063300,China)

机构地区：[1]沧州市人民医院骨科,沧州061000 [2]沧州市中心医院骨科,沧州061001 [3]唐山市丰南区医院骨科,唐山063300

出　　处：《西北药学杂志》2024年第4期64-69,共6页Northwest Pharmaceutical Journal

基　　金：河北省医学科技攻关计划项目(编号:20200332)。

摘　　要：目的探讨补骨脂素对骨肉瘤细胞侵袭、转移的抑制作用及可能的作用机制。方法用不同浓度补骨脂素作用于人骨肉瘤细胞(human osteosarcoma cells,MG-63),用CCK-8法检测细胞的存活情况,筛选最佳抑制浓度;用实时荧光定量法检测骨肉瘤组织与正常组织中微小RNA(microRNA,miR)-101的相对表达量。将对数生长期骨肉瘤细胞MG-63随机分为对照组、miR101模拟物阴性对照组(miR-NC组)、miR-101组、补骨脂素组和补骨脂素联合miR-101组。用四甲基偶氮唑盐[3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide,MTT]实验检测细胞增殖抑制率;用肿瘤细胞侵袭实验(Transwell)检测细胞侵袭和迁移能力;用实时荧光定量聚合酶链式反应(quantitative real time polymerase chain reaction,RT-PCR)检测肌磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)、蛋白激酶B(protein kinase B,Akt)mRNA的表达情况;用蛋白印迹法(Western blotting)检测磷酸化肌磷脂酰肌醇3-激酶(p-PI3K)、磷酸化蛋白激酶B(p-Akt)、基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)和基质金属蛋白酶9(matrix metalloproteinase-9,MMP-9)的表达情况。结果与对照组比较,miR-101组、补骨脂素组和补骨脂素联合miR-101组的细胞增殖抑制率升高,细胞侵袭数量、细胞迁移数量、PI3K和Akt mRNA,p-PI3K、p-Akt、MMP2和MMP9蛋白的表达水平均降低(P<0.05)。与miR-101组和补骨脂素组比较,补骨脂素联合miR-101组的细胞增殖抑制率升高,细胞侵袭数量、细胞迁移数量、PI3K和Akt mRNA,p-PI3K、p-Akt、MMP-2和MMP-9蛋白的表达水平均降低(P<0.05)。结论补骨脂素能够降低骨肉瘤细胞MG-63的增殖能力,并抑制其侵袭和迁移能力,其作用机制可能与调节miR-101水平、影响PI3K/Akt信号通路有关。Objective To investigate the inhibitory effect of psoralen on the invasion and metastasis of osteosarcoma cells and to explore its possible mechanism.Methods Psoralen with different concentrations acted on human osteosarcoma cells(MG-63).The cell survival was detected by CCK-8 method to screen the best inhibitory concentration.The relative expression of miR-101 in osteosarcoma and normal tissues was detected by real-time fluorescence quantitative method.The logarithmic growth MG-63 cells were randomly divided into control group,miR-NC group,miR-101 group,psoralen group,and psoralen combined with miR-101 group.The inhibition rate of cell proliferation was detected by 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide(MTT)assay,the ability of cell invasion and migration was detected by Transwell chamber test,and the mRNA expression of phosphatidylinositol 3-kinase(PI3K)and protein kinase B(Akt)was detected by quantitative real time polymerase chain reaction(RT-PCR).The expressions of phosphorylated myophosphatidylinositol 3-kinase(p-PI3K),phosphorylated protein kinase B(p-Akt),matrix metalloproteinase-2(MMP-2)and matrix metalloproteinase-9(MMP-9)were detected by Western blotting.Results Compared with the control group,the cell proliferation inhibition rate was increased,and the number of cell invasion,cell migration,PI3K and Akt mRNA,p-PI3K,p-Akt,MMP2 and MMP9 protein expression levels were decreased in miR-101 group,psoralen group,and psoralen combined with miR-101 group(P<0.05).Compared with miR-101 group and psoralen group,the cell proliferation inhibition rate was increased,and the number of cell invasion,cell migration,PI3K and Akt mRNA,p-PI3K,p-Akt,MMP-2 and MMP-9 protein expression levels were decreased in psoralen combined with miR-101 group(P<0.05).Conclusion Psoralen can reduce the proliferation of osteosarcoma cell line MG-63 and inhibit its invasion and migration.Its mechanism may be related to regulating the level of miR-101 and affecting PI3K/Akt signaling pathway.

关 键 词：补骨脂素 miR-101 人骨肉瘤细胞 肌磷脂酰肌醇3-激酶 蛋白激酶B 

分 类 号：R965[医药卫生—药理学]