染色质重塑蛋白MORC2通过调节ALDH1A3表达促进乳腺癌细胞的干性

发布时间：2018-04-22 01:40

本文选题：MORC + ALDHA　；参考：《中国癌症杂志》2017年03期

【摘要】：背景与目的:染色质重塑蛋白MORC家族CW型锌指结构蛋白2(microrchidia family CW-type zinc finger 2,MORC2)在DNA介导的基因转录及DNA损伤修复等基本生物学过程中发挥重要作用,但其对乳腺癌细胞生物学行为的影响目前尚无相关报道。乙醛脱氢酶(aldehyde dehydrogenase,ALDH)家族成员ALDH1A3(aldehyde dehydrogenase family 1 member A3,ALDH1A3)是一种公认的乳腺癌干细胞的标志物,但其在乳腺癌细胞中的调控机制目前尚不清楚。该研究拟探讨敲低MORC2基因对ALDH1A3表达以及对乳腺癌细胞干性的影响。方法:利用短发夹RNA(short hairpin RNA,sh RNA)介导的基因沉默方法构建稳定敲低MORC2基因的乳腺癌细胞系;利用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)和蛋白[质]印迹法(Western blot)分析敲低MORC2基因对ALDH1A3表达的影响;利用微球体形成实验和流式细胞荧光分选技术(fluorescence activated cell sorting,FACS)分析敲低MORC2对乳腺癌干细胞亚群的影响。结果:Western blot和RTFQ-PCR分析结果显示,敲低MORC2基因显著下调ALDH1A3蛋白及m RNA水平;微球体形成实验显示敲低MORC2基因显著抑制MCF-7细胞的成球能力;FACS分析显示敲低MORC2基因显著下调ALDH1A3阳性乳腺癌干细胞亚群,而对CD44+CD24-乳腺癌干细胞亚群没有明显影响。结论:MORC2通过调节ALDH1A3的表达促进乳腺癌干细胞表型。
[Abstract]:Background & AIM: the CW zinc finger protein 2(microrchidia family CW-type zinc finger 2 Moc2 of chromatin remodeling protein MORC family plays an important role in DNA mediated gene transcription and DNA damage repair. However, there is no related report on the effect on the biological behavior of breast cancer cells. ALDH1A3(aldehyde dehydrogenase family 1 member A3A3 is a recognized marker of breast cancer stem cells, but its regulatory mechanism in breast cancer cells is still unclear. The aim of this study was to investigate the effects of MORC2 knockout on ALDH1A3 expression and dryness of breast cancer cells. Methods: a breast cancer cell line with stable knockout of MORC2 gene was constructed by short hairpin RNA(short hairpin RNA-mediated gene silencing. Real-time fluorescent quantitative polymerase chain reactionation (RTFQ-PCR) and Western blotting were used to analyze the effect of MORC2 gene knockout on ALDH1A3 expression. Microsphere formation test and flow cytometry (FCM) were used to analyze the effect of low MORC2 on breast cancer stem cell subsets. Results the results of Western blot and RTFQ-PCR analysis showed that knockdown of MORC2 gene significantly down-regulated the level of ALDH1A3 protein and m RNA. Microsphere formation assay showed that knockout MORC2 gene significantly inhibited the spheroidization of MCF-7 cells. FACS analysis showed that knockout MORC2 gene significantly down-regulated ALDH1A3 positive breast cancer stem cell subsets, but had no significant effect on CD44 CD24- breast cancer stem cell subsets. Conclusion the expression of ALDH1A3 in breast cancer stem cells can be promoted by adjusting the expression of ALDH1A3 in the breast cancer stem cell phenotype.
【作者单位】：复旦大学生物医学研究院及肿瘤研究所;
【基金】：国家自然科学基金(81372847)
【分类号】：R737.9

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