下调MTHFD2对前列腺癌PC3细胞的侵袭和转移的作用及影响机制

    Down-regulation of MTHFD2 inhibits invasion and metastasis in prostate cancer PC3 cells

    • 摘要:
      目的观察线粒体亚甲基四氢叶酸脱氢酶2(mitochondrial methylenetetrahydrofolate dehydrogenase 2, MTHFD2)基因表达对人前列腺癌PC3细胞侵袭和转移能力的影响。
      方法人前列腺癌PC3细胞分为3组: 空白对照组(ctrl组)、阴性对照组(si-NC组)及转染siRNA-MTHFD2组(si-MTHFD2组)。采用qRT-PCR法检测人正常前列腺上皮细胞RWPE-1以及前列腺癌PC3细胞中MTHFD2的mRNA表达,通过流式细胞术检测MTHFD2对PC3细胞凋亡的影响,通过划痕实验检测细胞迁移能力的变化,通过Transwell实验检测细胞侵袭能力的变化,采用Western blotting检测E-cadherin、Vimentin和N-cadherin蛋白表达变化。
      结果前列腺癌PC3细胞较正常前列腺上皮细胞中MTHFD2表达水平升高(P < 0.05),si-MTHFD2组MTHFD2 mRNA表达水平较ctrl组和si-NC组降低(P < 0.05);si-MTHFD2组细胞迁移和侵袭能力较ctrl组和si-NC组降低(P < 0.05);si-MTHFD2组E-candherin蛋白表达水平较ctrl组和si-NC组升高(P < 0.05),Vimentin和N-cadherin蛋白表达水平较ctrl组和si-NC组降低(P < 0.05)。
      结论下调MTHFD2表达可增加前列腺癌细胞凋亡能力,抑制前列腺癌细胞迁移和侵袭能力。

       

      Abstract:
      ObjectiveTo investigate the effect of mitochondrial methylenetetrahydrofolate dehydrogenase 2 (MTHFD2) on cell invasion and metastasis in prostate cancer PC3 cells.
      MethodsThe prostate cancer PC3 cells were divided into 3 groups: blank control group (ctrl group), negative control group (si-NC group) and transfected siRNA-MTHFD2 group (si-MTHFD2 group).The mRNA expression of MTHFD2 in human normal prostate epithelial cells RWPE-1 and prostate cancer PC3 cells was detected by qRT-PCR.The apoptosis rate in different groups was detected by flow cytometry, the changes of cell migration ability were detected by scratch test, the changes of cell invasion ability were detected by Transwell test, and the changes of E-cadherin, Vimentin and N-cadherin protein expressions were detected by Western blotting.
      ResultsThe expression level of MTHFD2 in prostate cancer PC3 cells was significantly higher than that in normal prostate epithelial cells(P < 0.05), and the mRNA expression level of MTHFD2 in si-MTHFD2 group was significantly lower than that in ctrl and si-NC groups(P < 0.05).The results of cell scratch and invasion assay showed that the migration and invasion abilities in si-MTHFD2 group was significantly lower than in ctrl and si-NC groups(P < 0.05).E-candherin protein expression level in si-MTHFD2 group was significantly higher than that in ctrl and si-NC groups(P < 0.05), and the protein expression levels of vimentin and N-cadherin were significantly lower than those in ctrl and si-NC groups.
      ConclusionsDown-regulation of MTHFD2 expression can increase the apoptosis of prostate cancer cells and inhibit the migration and invasion of prostate cancer cells.

       

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