FBXO11基因对肺腺癌A549细胞迁移的影响

发布时间：2018-05-07 12:23

本文选题：肺腺癌 + FBXO11　；参考：《安徽医科大学》2017年硕士论文

【摘要】：目的:研究泛素连接酶复合体(Skp1 Cul1 F-box-protein,SCF)特异性亚基FBXO11的表达对肺腺癌A549细胞迁移的影响。方法:将培养的肺腺癌A549细胞株分为三组:空白组(Blank组)、阴性对照RNA组(siRNA-NC组)、FBXO11干扰组(siRNA-FBXO11组),应用Li-pofectamine 2000将siRNA-FBXO11、siRNA-NC组分别转染至siRNA-FBXO11组和siRNA-NC组的A549细胞中。(1)应用试剂Trizol提取A549细胞中总的RNA,RT-PCR半定量的方法检测FBXO11的mRNA水平;(2)免疫印迹法(Western-Blot)法检测FBXO11和Snail的蛋白水平;(3)划痕修复实验检测A549细胞的迁移能力。结果:(1)RT-PCR结果示,Blank组、siRNA-NC组和siRNA-FBXO11组之间比较有差异,且差异具有统计学意义(P0.05)。与Blank组、siRNA-NC组比较,siRNA-FBXO11组FBXO11的mRNA水平显著降低(P0.05),而Blank组与siRNA-NC组比较差异无统计学意义(P0.05)。(2)Western-Blot结果示:FBXO11蛋白水平比较,三组之间有差异,且差异具有统计学意义(P0.05)。与Blank组、siRNA-NC组比较,siRNA-FBXO11组FBXO11的蛋白水平显著降低(P0.05),而Blank组与siRNA-NC组比较差异无统计学意义(P0.05)。Snail蛋白水平比较,三组之间有差异,且差异有统计学意义(P0.05)。与Blank组、siRNA-NC组比较,siRNA-FBXO11组的snail蛋白表达水平显著增加(P0.05),而Blank组与siRNA-NC组比较无显著性差异(P0.05)。(3)划痕实验结果显示:比较三组之间的迁移能力有差异,且差异具有统计学意义(P0.05)。与Blank组、siRNA-NC组比较,siRNA-FBXO11组的细胞迁移能力增强(P0.05)。Blank组与siRNA-NC组比较差异无统计学意义(P0.05)。结论:FBXO11基因的表达能抑制肺腺癌A549细胞的迁移能力,这可能与Snail在细胞内的调控有关。
[Abstract]:Aim: to study the effect of the expression of the specific subunit FBXO11 of Skp1 Cul1 and F-box-protein Cul1 on the migration of lung adenocarcinoma A549 cells. Methods: the cultured lung adenocarcinoma A549 cell line was divided into three groups: blank group (Blank group), negative control group (RNA group), siRNA-NC group (FBXO11 interference group), siRNA-FBXO11 interference group (FBXO11 group). SiRNA-FBXO11siRNA-NC group was transfected into A549 cells of siRNA-FBXO11 group and siRNA-NC group by Li-pofectamine 2000. Trizol reagent was used to extract Trizol. The mRNA level of FBXO11 in A549 cells was detected by semi-quantitative RT-PCR. Western blotting was used to detect the protein level of FBXO11 and Snail in A549 cells. The migration ability of A549 cells was detected by scratch repair assay. Results the results of RT-PCR showed that there was a significant difference between the siRNA-FBXO11 group and the control group, and the difference was statistically significant (P 0.05). Compared with the Blank group, the mRNA level of the FBXO11 in the siRNA-FBXO11 group was significantly lower than that in the siRNA-NC group, but there was no significant difference between the Blank group and the siRNA-NC group. The results of Western-Blot showed that there was a significant difference among the three groups, and the difference was statistically significant. Compared with Blank group, FBXO11 protein level of siRNA-FBXO11 group was significantly lower than that of siRNA-NC group, but there was no significant difference between Blank group and siRNA-NC group. There was significant difference among the three groups, and the difference was statistically significant (P 0.05). Compared with the Blank group, the expression of snail protein in the siRNA-FBXO11 group was significantly higher than that in the siRNA-NC group, but there was no significant difference between the Blank group and the siRNA-NC group. The results of scratch test showed that the migration ability of the three groups was different and the difference was statistically significant (P 0.05). Compared with Blank group, siRNA-FBXO11 group could enhance cell migration ability. There was no significant difference between siRNA-NC group and siRNA-NC group (P 0.05). Conclusion the expression of FBXO11 gene can inhibit the migration of lung adenocarcinoma A549 cells, which may be related to the regulation of Snail in the cells.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R734.2

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