宫颈癌细胞中ΔNp63α对白血病抑制因子表达水平的影响及其机制

徐菲; 钱莉莉; 王慧妍; 孙正伟; 程勇; 赵卫东; 肖卫华; 周颖

doi:10.3971/j.issn.1000-8578.2016.07.001

宫颈癌细胞中ΔNp63α对白血病抑制因子表达水平的影响及其机制

Effect ofΔNp63αon Leukemia Inhibitory Factor Expression in Cervical Cancer Cells and Related Mechanism

摘要

摘要:
目的探讨ΔNp63α对白血病抑制因子表达水平的影响及其机制。
方法通过稳定转染构建稳定过表达ΔNp63α的SiHa细胞株(SiHa/ΔNp63α)；通过瞬时转染沉默ME-180细胞中ΔNp63α的表达水平；并通过Western blot检测两组细胞中ΔNp63α、LIF、STAT3的蛋白水平，通过qRT-PCR检测两组细胞中ΔNp63α、LIF、STAT3、LncRNA ENST00000447565的转录水平。
结果 (1)ΔNp63α过表达的SiHa细胞中，LIF的mRNA及蛋白水平均明显下调，其调控的下游基因STAT3的mRNA表达水平也显著下降；(2)ΔNp63α过表达的SiHa细胞中，LncRNA ENST00000447565的转录水平显著下调，该LncRNA与LIF基因均在22号染色体q12.2，且基因组位置接近；(3)ΔNp63α沉默的ME-180细胞中，LIF的表达量明显上调。
结论 ΔNp63α可以抑制LIF及下游基因STAT3的表达水平，其机制可能与ΔNp63α对LncRNA ENST00000447565的表达抑制有关。

Abstract:
Objective To explore the effect ofΔNp63αon the expression level of leukemia inhibitory factor (LIF) and its mechanism.
Methods Stable SiHa cells with overexpressedΔNp63α(SiHa/ΔNp63α) were established by stable transfection, andΔNp63αexpression was transiently silenced in ME-180 cell lines(ME-180/ΔNp63αsiRNA). The protein levels ofΔNp63α, LIF and STAT3 were detected by Western blot, while the mRNA levels ofΔNp63α, LIF, STAT3 and LncRNA ENST00000447565 were detected by qRT-PCR.
Results (1) The mRNA and protein levels of LIF and its downstream signaling factor STAT3 mRNA was significantly reduced in SiHa/ΔNp63αcells; (2) The RNA levels of LncRNA ENST00000447565 were significantly reduced in SiHa/ΔNp63αcells, ENST00000447565 and LIF were both located in chromosome 22 q12.2, within the same loci; (3) The mRNA level of LIF was significantly increased in ME-180 cells with silencedΔNp63α.
Conclusion The expressions of LIF and its downstream signaling factor STAT3 are inhibited byΔNp63αexpression, which might be associated with the down-regulation of LncRNA ENST00000447565.

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