FGL1调控非小细胞肺癌PC9/GR细胞迁移和侵袭机制的研究

Mechanism of FGL1 regulates migration andinvasion of PC9/GR cells of non-small cell lung cancer

目的研究FGL1在PC9细胞和PC9/GR细胞中FGL1的表达差异,探讨FGL1对肺腺癌细胞PC9/GR细胞迁移和侵袭的影响机制。方法采用siRNA法沉默PC9/GR细胞FGL1,RT-qPCR和Western blot法确定干扰效果;划痕实验、Transwell小室实验检测细胞迁移侵袭能力;Western blot法检测细胞中的Zeb1、Vimentin和E-cadherin表达量。结果 RT-qPCR和Western blot显示PC9/GR细胞中FGL1表达水平明显高于PC9细胞;CCK-8实验显示PC9/GR细胞IC50明显高于PC9细胞,敲低FGL1后IC50明显降低;划痕实验、Traswell小室侵袭迁移实验显示,干扰PC9/GR细胞中FGL1后细胞伤口愈合能力、迁移和侵袭能力明显减弱;在相同剂量吉非替尼药物作用下,干扰FGL1后PC9/GR细胞伤口愈合能力、细胞迁移和侵袭能力减弱更加明显;Western blot提示干扰FGL1可明显降低PC9/GR细胞Zeb1、Vimentin表达而增加E-cadherin表达。结论 FGL1在PC9/GR细胞中表达显著升高,干扰FGL1可通过抑制EMT过程,抑制PC9/GR细胞的侵袭和迁移。

Aim To explore the expression of FGL1 in PC9 cells and PC9/GR cells and to investigate the mechanism of FGL1 regulating migration and invasion of PC9/GR cells.Methods FGL1 of PC9/GR cells was silenced by siRNA The interference effect was proved by RT-qPCR and Western blot.The IC 50 of PC9 cells and PC9/GR cells was detected by CCK-8 method.Cell migration and invasion were detected by Transwell assay and Scratch test.The expressions of E-cadherin,Zeb1 and Vimentin were measured by Western blot.Results RT-qPCR and Western blot showed that the expression of FGL1 in PC9/GR cells was significantly higher than that in PC9 cells.CCK-8 showed that IC 50 of PC9/GR cells was significantly higher than that of PC9 cells,and evidently decreased after FGL1 knockdown.The scratch test and Traswell assay showed that the wound healing ability,migration and invasion ability of PC9/GR cells were significantly weakened when FGL1 was reduced.And under the same dose of gefitinib,the wound healing ability,migration and invasion ability of PC9/GR cells were more significantly weakened after FGL1 was interfered.Western blot showed that silencing the expression of FGL1 could significantly reduce the expression of Zeb1 and vimentin in PC9/GR cells but contrarily increase E-cadherin.Conclusions FGL1 significantly increases in PC9/GR cells.Inhibiting the expression of FGL1 can significantly inhibit the invasion and metastasis of PC9/GR cell by suppressing EMT.