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乳腺癌MCF-7细胞中LncRNA XIST的表达及功能研究

Expression and Functional of LncRNA XIST in MCF-7 Breast Cancer Cells

  • 摘要:
    目的 探讨lncRNA XIST在乳腺癌MCF-7细胞中的表达及其对MCF-7细胞功能的影响。
    方法 qRT-PCR检测人正常乳腺上皮细胞MCF-10A及人乳腺癌MCF-7细胞中XIST的表达水平;利用瞬时转染技术在MCF-7细胞中过表达XIST,MTT法和Transwell小室法检测XIST过表达后MCF-7细胞增殖、迁移和侵袭能力的变化情况;生物信息学预测并用qRT-PCR检测与XIST有潜在结合位点的miRNA表达变化情况;并在MCF-7细胞中瞬时转染miR-130b-3p Inhibitor,qRT-PCR检测细胞中XIST的表达水平。
    结果 乳腺癌MCF-7细胞中XIST低表达(P < 0.001),miR-130b-3p高表达(P < 0.001)。XIST过表达可显著抑制MCF-7细胞的增殖(P < 0.001)、促进MCF-7细胞的迁移和侵袭,而且XIST过表达后MCF-7细胞中miR-130b-3p表达水平显著降低(P < 0.01),miR-130b-3p低表达后MCF-7细胞中XIST表达水平显著增加(P < 0.001)。
    结论 乳腺癌MCF-7细胞中XIST和miR-130b-3p的表达呈负相关,且XIST过表达可显著抑制MCF-7细胞的增殖、促进迁移和侵袭。

     

    Abstract:
    Objective To investigate the expression and function of lncRNA XIST in MCF-7 breast cancer cells.
    Methods The expression of XIST in MCF 10A human normal breast epithelial cells and MCF-7 human breast cancer cells were detected by qRT-PCR. Transient transfection technique was used to overexpress XIST in MCF-7 cells. MTT assay and Transwell assay were used to detect the changes of proliferation, migration and invasion of MCF-7 cells after XIST overexpression in MCF-7 cells. Bioinformatics and qRT-PCR were used to predict and detect the changes of miRNA expression which have potential binding sites of XIST. qRT-PCR was used to detect the expression of XIST in MCF-7 cells which was transfected instantaneously by miR-130b-3p inhibitor.
    Results XIST expression was decreased in MCF-7 cells, while miR-130b-3p expression was increased, compared with those in MCF-10A cells (both P < 0.001). Overexpression of XIST significantly inhibited the proliferation of MCF-7 cells (P < 0.001), promoted cell migration and invasion. And the expression level of miR-130b-3p in MCF-7 cells was reduced significantly after overexpression of XIST (P < 0.01), while the expression level of XIST in MCF-7 cells was increased significantly after down-expression of miR-130b-3p (P < 0.001).
    Conclusion The expression levels of XIST and miR-130b-3p in breast cancer MCF-7 cells are negatively correlated, and the overexpression of XIST could significantly inhibit the proliferation, and promote the migration and invasion of MCF-7 cells.

     

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