槲皮素联合LAP3基因沉默对非小细胞肺癌A549细胞功能的影响及其机制-现代医学

槲皮素联合LAP3基因沉默对非小细胞肺癌A549细胞功能的影响及其机制

单位：1. 枣庄矿业集团中心医院胸外科, 山东枣庄 277000;
2. 枣庄矿业集团中心医院重症医学科, 山东枣庄 277000

分类号：Q291

出版年·卷·期（页码）：2020·39·第十二期（1577-1582）

摘要：

目的： 探讨槲皮素联合亮氨酸氨肽酶3（LAP3）基因沉默对非小细胞肺癌A549细胞增殖、侵袭和迁移的影响及其机制。方法： 以不同浓度（0、5、10、20、40和80 μmol·L^-1）的槲皮素处理A549细胞，筛选槲皮素的作用浓度。在A549细胞中转染LAP3 siRNA，实验分为阴性组、槲皮素、LAP3干扰组和槲皮素+LAP3干扰组，采用噻唑蓝（MTT）法、克隆形成实验和Transwell小室分别检测A549细胞的增殖、侵袭、迁移能力，免疫印迹法检测A549细胞中LAP3、上皮型钙黏蛋白、N-钙黏附素、波形蛋白的表达。结果： 10、20、40和80 μmol·L^-1槲皮素处理后A549细胞存活率逐渐降低，且与0 μmol·L^-1比较，差异均有统计学意义（P<0.05），槲皮素的半抑制浓度为46.16 μmol·L^-1。与阴性组比较，LAP3干扰组、槲皮素、槲皮素+LAP3干扰组细胞存活率、克隆形成率、侵袭细胞数、迁移细胞数以及LAP3、N-钙黏附素、波形蛋白的表达水平均显著降低（均P<0.05），而细胞中上皮型钙黏蛋白的表达水平显著升高（P<0.05），且槲皮素+LAP3干扰组中的上述变化较LAP3干扰组或槲皮素组更为显著（P<0.05）。结论： 槲皮素联合LAP3基因沉默可协同抑制非小细胞肺癌A549细胞增殖，并可通过共同减弱细胞上皮间质转化进程协同抑制细胞侵袭和迁移。

Objective: To investigate the effects of quercetin combined with leucine aminopeptidase 3(LAP3)gene silencing on proliferation, invasion and migration of non-small cell lung cancer A549 cells and its mechanism. Methods: A549 cells were treated with quercetin at different concentrations (0, 5, 10, 20, 40 and 80 μmol·L^-1) to screen the concentration of quercetin. LAP3 siRNA was transfected into A549 cells, and the experiment was divided into negative group, quercetin group, LAP3 interference group and quercetin+LAP3 interference group. Thiazolyl blue (MTT) method, clone formation experiment and Transwell chamber were used to detect the proliferation, invasion and migration ability of A549 cells respectively. Western blotting was used to detect the expression of LAP3, E-cadherin, N-cadherin and Vimentin protein in A549 cells.Results: After 10, 20, 40 and 80 μmol·L^-1 quercetin treatment, the survival rate of A549 cells gradually decreased, and compared with 0 μmol·L^-1, the difference was statistically significant (P<0.05). The half-inhibitory concentration of quercetinwas 46.16 μmol·L^-1. Compared with the negative group, LAP3 interference group, quercetin group, quercetin+LAP3 interference group cell survival rate, clone formation rate, number of invasion cells, number of migration cells, LAP3, N-cadherin,Vimentin protein expression were all significantly reduced (all P<0.05), while the expression levels of E-cadherin protein in cells were significantly higher (P<0.05). The above changes in quercetin+LAP3 interference group were more significant than those in LAP3 interference group or quercetin group (P<0.05).Conclusion: Quercetin combined with LAP3 gene silencing can synergistically inhibit the proliferation of non-small cell lung cancer A549 cells, and synergistically inhibit cell invasion and migration by attenuating the process of cell epithelial-mesenchymal transition.

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