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TIP30基因对大肠癌细胞HCT116生物学特性的影响

Effect of TIP30 on Biological Characters of HCT116 Cells

  • 摘要: 目的 通过构建基因真核表达载体分析TIP30对大肠癌细胞HCT116生物学特性的影响,为TIP30在大肠癌基因治疗中的应用提供依据。方法构建pCMV4-flag-TIP30真核表达载体并转染HCT116细胞,RT-PCR和Western blot检测TIP30基因表达,体外侵袭实验检测细胞侵袭能力,软琼脂实验检测细胞成瘤性。结果成功构建稳定表达TIP30的HCT116细胞模型,转染TIP30 的HCT116细胞增殖受抑,侵袭能力及克隆形成能力均减弱。结论大肠癌HCT116细胞TIP30过表达不仅能抑制其生长、诱导其凋亡,并能降低其侵袭、迁移能力,为TIP30基因治疗提供依据。

     

    Abstract: Objective To study the association of TIP30 and biological characters of HCT116 colorectal cancer cells. Methods The eukaryotic expression vector of TIP30 gene was established and transfected into HCT116 cells by liposome-mediated transfection.The expressions of TIP30 gene mRNA and protein were detected by RT-PCR and Western blot.The invasive ability was analyzed by Matrigel invasion assay.The colony formation assay was used to detect cell growth inhibition. Results The stable transfectant of HCT116/pCMV4-flag-TIP30 cell was established successfully,the proliferation of HCT116/pCMV4-flag-TIP30 cell was inhibited,the invasive ability and colony formation ability were also decreased compared to control cell. Conclusion The overexpression of TIP30 gene in HCT116 cells not only results in inhibiting cell growth and cell apoptosis,but also reducing cell migration and invasion,which provides evidence for TIP30 gene therapy.

     

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