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NRP-1过表达对乳腺癌细胞MDA-MB-231、SK-BR-3生物学行为的影响

Effects of NRP-1 Overexpression on Biological Behaviors of Breast Cancer Cell Lines MDA-MB-231 and SK-BR-3

  • 摘要:
    目的  通过上调乳腺癌细胞MDA-MB-231、SK-BR-3中NRP-1的表达,观察NRP-1对细胞增殖、凋亡、迁移及侵袭能力的影响。
    方法  构建pcDNA3.1-NRP-1表达载体,脂质体介导NRP-1表达质粒转染MDA-MB-231、SK-BR-3细胞,用G418筛选出稳定转染的乳腺癌细胞株。利用RT-qPCR、Western blot法分别检测NRP-1基因mRNA及其蛋白表达;CCK-8法、AnnexinⅤ-APC/7-AAD法、Transwell小室分别检测转染细胞增殖率、凋亡率及侵袭、迁移能力。
    结果  成功构建pcDNA3.1-NRP-1表达载体,转染MDA-MB-231、SK-BR-3细胞并筛选稳定表达系。与对照组相比,过表达组细胞的NRP-1 mRNA及蛋白表达水平明显升高(均P<0.05);NRP-1过表达组的细胞较对照组增殖率增加、凋亡率降低、侵袭及迁移能力增强(均P<0.05)。
    结论  NRP-1在乳腺癌发展、浸润、转移中起着一定的作用,它可促进乳腺癌细胞增殖、迁移和侵袭,抑制其凋亡。

     

    Abstract:
    Objective  To upregulate neuropilin-1(NRP-1) expression in breast cancer cell lines MDA-MB-231 and SK-BR-3, and to investigate their effects on cells proliferation, apoptosis, invasion and migration.
    Methods  PcDNA3.1-NRP-1 expression vector was constructed and transfected into MDA-MB-231 and SK-BR-3 cell lines by liposome, then stably expressing cell lines were screened by different concentrations of G418. The expression levels of NRP-1 mRNA and protein were detected by RT-qPCR and Western blot. We used CCK-8 assay, AnnexinⅤ-APC/7-AAD assay, Transwell assay to detect the change of cells proliferation, apoptosis, invasion and migration.
    Results  pcDNA3.1-NRP-1 expression vector was successfully constructed and transfected into MDA-MB-231 and SK-BR-3 cell lines, the stably expressing cell lines were screened. NRP-1 expression was up-regulated significantly at both mRNA and protein levels, compared with control group(P<0.05). CCK-8 assay and Transwell assay showed that NRP-1 promoted the proliferation, invasion and migration of MDA-MB-231 and SK-BR-3 cell lines(all P<0.05). AnnexinⅤ-APC/7-AAD assay showed that NRP-1 inhibited the apoptosis of the two cell lines(P<0.05).
    Conclusion  NRP-1 plays a specific role in the development, invasion and migration of breast cancer. It can promote the proliferation, invasion and migration, and inhibit the apoptosis of breast cancer cells.

     

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