非洲爪蟾驱动蛋白样蛋白2靶蛋白对非小细胞肺癌细胞迁移侵袭的影响及其机制  

Effect and mechanism of TPX2 on the migration and invasion of non-small cell lung cancer cells

作　　者：刘雷 施民新[1] 陆海敏[1] 王伟[1] LIU Lei;SHI Minxin;LU Haimin;WANG Wei(Department of Thoracic Surgery,The Affiliated Tumor Hospital of Nantong University,Nantong,Jiangsu 226300,China)

机构地区：[1]南通大学附属肿瘤医院胸外科,江苏南通226300

出　　处：《安徽医药》2021年第12期2453-2458,共6页Anhui Medical and Pharmaceutical Journal

基　　金：南通卫生青年基金(WKZL2018049)。

摘　　要：目的研究非洲爪蟾驱动蛋白样蛋白2靶蛋白(TPX2)基因对非小细胞肺癌细胞迁移侵袭的影响及其机制。方法2018年10月至2019年8月,将非小细胞肺癌A549、H1299细胞株分为三组:空白对照组(未经转染的细胞);阴性对照组[转染非特异性的小干扰RNA(siRNA)的细胞];siRNA-TPX2组(转染TPX2特异性的siRNA的细胞)。采用实时荧光定量逆转录聚合酶链反应(qRT-PCR)和蛋白质印迹法(Western blotting)方法对转染小干扰RNA(siRNA)前后的非小细胞肺癌A549、H1299细胞株进行鉴定。人胆囊收缩素/缩胆囊素八肽(CCK-8)增殖实验检测细胞增殖能力。Tanswell小室迁移实验检测细胞迁移和侵袭的影响。蛋白质印迹法检测磷脂酰肌醇3激酶(PI3K)-蛋白激酶B(Akt)信号通路关键分子及基质金属蛋白酶(MMPs)表达的变化。结果siRNA-TPX2组中TPX2 mRNA和蛋白的相对表达水平显著低于阴性对照组及空白对照组(P<0.05)。与阴性对照组相比,下调TPX2可抑制A549细胞的增殖[24 h(0.71±0.02)比(0.62±0.03);48 h(1.62±0.03)比(0.69±0.02);72 h(2.08±0.03)比(0.95±0.02)]、迁移[(91.18±13.97)个比(49.95±12.15)个]、侵袭[(165.46±19.19)个比(93.37±13.54)个](P<0.05)。与阴性对照组相比,下调TPX2可抑制H1299细胞的增殖[24 h(0.81±0.03)比(0.65±0.02);48 h(1.73±0.02)比(0.78±0.01);72 h(2.18±0.03)比(1.16±0.02)]、迁移[(74.31±13.86)个比(38.76±9.05)个]、侵袭[(158.46±19.58)个比(88.47±11.23)个](P<0.05)。TPX2表达下调能显著降低非小细胞肺癌A549、H1299细胞中PI3K、磷酸化蛋白激酶B(p-Akt)、基质金属蛋白酶9(MMP9)和基质金属蛋白酶12(MMP12)蛋白的表达(P<0.05)。激动剂胰岛素生长样因子1(IGF-1)能完全削弱TPX2-siRNA抑制A549、H1299细胞中PI3K/Akt信号通路活性及其下游MMP9和MMP12的表达(P<0.05)。结论TPX2通过调控PI3K/Akt信号通路活性及其下游MMP9和MMP12的表达促进非小细胞肺癌细胞侵袭转移。Objective To investigate the effects of Xenopus kinesin-like protein 2 target protein(TPX2)expression on tumor migration and invasion of Non-small cell lung cancer cell and its underlying mechanism.Methods From October 2018 to August 2019,A549 and H1299 cell lines of non-small cell lung cancer(NSCLC)were assigned into three groups:blank control group(untransfected cells),negative control group(transfected with non-specific small interfering RNA(siRNA)cells],and siRNA-TPX2 group(cells transfected with TPX2 specific siRNA).Real-time quantitative PCR and Western blotting were used to identify the A549 and H1299 cell lines before and after transfection with siRNA.The proliferation of cells was evaluated by CCK-8 proliferation experiment.The migration and invasion of cells were assessed by transwell chambers experiment.The changes of phosphatidylinositol 3-kinase(PI3K)-protein kinase B(Akt)pathway-realted proteins and Matrix Metalloproteinases(MMPs)protein expressions were measured by Western blotting.Results The levels of TPX2 mRNA and protein in siRNA-TPX2 group were significantly lower than those in negative control group and blank group(P<0.05).Compared with negative control group,down-regulation of TPX2 could inhibit the proliferation of A549 cells[24 h(0.71±0.02)vs.(0.62±0.03);48 h(1.62±0.03)vs.(0.69±0.02);72 h(2.08±0.03)vs.(0.95±0.02)],migration[(91.18±13.97)vs.(49.95±12.15)],invasion[(165.46±19.19)vs.(93.37±13.54)](P<0.05).Compared with negative control group,down-regulation of TPX2 could inhibit the proliferation of H1299 cells[24 h(0.81±0.03)vs.(0.65±0.02);48 h(1.73±0.02)vs.(0.78±0.01);72 h(2.18±0.03)vs.(1.16±0.02)],migration[(74.31±13.86)vs.(38.76±9.05)],and invasion[(158.46±19.58)vs.(88.47±11.23)](P<0.05).Down-regulation of TPX2 markedly decreased the expressions of PI3K,p-Akt,MMP9 and MMP12 proteins in A549 and H1299 cells of NSCLC(P<0.05).The agonist insulin growth-like factor 1(IGF-1)could attenuate the inhibition of PI3K/Akt signaling pathway activity and the downstream expression o

关 键 词：癌 非小细胞肺 基质金属蛋白酶9 基质金属蛋白酶12 非洲爪蟾驱动蛋白样蛋白2靶蛋白基因 侵袭 PI3K-AKT信号通路 

分 类 号：R734.2[医药卫生—肿瘤]