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宫颈癌细胞中ΔNp63α的MicroRNA表达谱及其对hsa-let-7b-3p的调控机制

MicroRNA Expression Profile of ΔNp63α in Cervical Cancer Cells and Its Regulation Mechanism on hsa-let-7b-3p

  • 摘要:
    目的 在宫颈鳞癌细胞中筛选出ΔNp63α显著调控的miRNAs,并探讨其对宫颈癌细胞功能的影响。
    方法 通过miRNA芯片技术筛选出ΔNp63α过表达的SiHa细胞株(SiHa/p63)中差异明显的miRNAs,并进行qRT-PCR验证;通过qRT-PCR在沉默ΔNp63α的ME-180细胞株(ME-180/Shp63)中检测差异表达的miRNAs,并选择差异较显著的hsa-let-7b-3p作为研究对象;向SiHa细胞中转染hsa-let-7b-3p刺激剂,通过生长曲线、Western blot以及流式细胞仪检测hsa-let-7b-3p对SiHa细胞增殖及凋亡的影响。
    结果 (1)在ΔNp63α过表达的SiHa细胞株中,筛选出10个变化较显著的miRNAs;(2)在ΔNp63α沉默的ME-180细胞中,筛选出5个与上述变化趋势一致的miRNAs;(3)hsa-let-7b-3p过表达抑制SiHa细胞的增殖;(4)hsa-let-7b-3p过表达促进SiHa细胞的凋亡。
    结论 宫颈癌细胞株中,miRNA hsa-let-7b-3p的表达与ΔNp63α的表达呈正相关,且可以有效抑制细胞增殖,其作用机制可能与诱导细胞周期阻滞和促进细胞凋亡相关。

     

    Abstract:
    Objective To screen out the miRNAs regulated by ΔNp63α in squamous cell carcinoma of the cervix and investigate their effect on the function of cervical cancer cells.
    Methods miRNA microarray was used to screen the miRNAs with significant difference in SiHa cells with overexpressed ΔNp63α and verified by qRT-PCR. The miRNAs with differential expression were validated by qRT-PCR in ME-180 cells with silenced ΔNp63α and the hsa-let-7b-3p was selected as the study object. The effect of hsa-let-7b-3p on the proliferation and apoptosis of SiHa cells was detected by growth curve, Western blot and flow cytometry after the transfection of hsa-let-7b-3p mimic into SiHa cells.
    Results (1) In SiHa cells with ΔNp63α overexpression, ten miRNAs consistent with the trend of qRT-PCR were screened out from the miRNAs with significant differences in the results of the microarray. (2) Five miRNAs that were consistent with the above change trends were screened out in ME-180 cells with silenced ΔNp63α. (3) Overexpression of hsa-let-7b-3p inhibited the proliferation of SiHa cells. (4) Overexpression of hsa-let-7b-3p promoted the apoptosis of SiHa cells.
    Conclusion In the cervical cancer cell line, the expression of miRNA hsa-let-7b-3p is positively correlated with the expression of ΔNp63α, and it can inhibit cell proliferation effectively. The mechanism may be related to the induction of cell cycle arrest and the promotion of apoptosis.

     

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