| 刘雷,施民新,陆海敏,等.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[J].安徽医药,2021,25(12):2453-2458. |
| 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 |
| |
| DOI：10.3969/j.issn.1009-6469.2021.12.028 |
| 中文关键词: 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 |
| 英文关键词: Carcinoma, non-small-cell lung Matrix metalloproteinase 9 Matrix metalloproteinase 12 TPX2 gene Nvasion PI3K/Akt signaling |
| 基金项目:南通卫生青年基金( WKZL2018049) |
|
| 摘要点击次数: 1148 |
| 全文下载次数: 376 |
| 中文摘要: |
| 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。 |
| 英文摘要: |
| Objective To investigate the effects of Xenopus kinesin-like protein 2 target protein (TPX2) expression on tumor migra. tion 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 (untransfectedcells), negative control group (transfected with non-specific small interfering RNA (siRNA) cells], and siRNA-TPX2 group (cells trans. fected with TPX2 specific siRNA). Real-time quantitative PCR and Western blotting were used to identify the A549 and H1299 celllines before and after transfection with siRNA. The proliferation of cells was evaluated by CCK-8 proliferation experiment. The migra.tion 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 down.stream expression of MMP9 and MMP12 in A549 and H1299 cells by siRNA-TPX2 (P<0.05).Conclusion TPX2 promotes invasionand metastasis of NSCLC by regulating PI3K/Akt signaling pathway activity and its downstream MMP9 and MMP12 expression. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
