Magnetofection based on superparamagnetic iron oxide nanoparticle-mediated low lncRNA HOTAIR expression decreases the proliferation and invasion of glioma stem cells.
Magnetofection based on superparamagnetic iron oxide nanoparticle-mediated low lncRNA HOTAIR expression decreases the proliferation and invasion of glioma stem cells.
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基于超顺磁性氧化铁纳米颗粒介导的低lncRNA HOTAIR表达的磁转染降低了胶质瘤干细胞的增殖和侵袭
DOI：
10.3892/ijo.2016.3571
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发表时间：
2016-08
影响因子：
5.2
通讯作者：
Wu Y
中科院分区：
文献类型：
--
作者：
Fang K;Liu P;Dong S;Guo Y;Cui X;Zhu X;Li X;Jiang L;Liu T;Wu Y
胶质瘤干细胞（GSC）是胶质瘤细胞的一个特殊亚群，是肿瘤对治疗敏感性和肿瘤复发可能性的关键。因此，确定抑制GSC生长的新策略对于开发多形性胶质母细胞瘤（GBM）的新疗法非常重要。本研究分离培养了CD 133+人胶质瘤干细胞。磁性纳米颗粒用于介导靶向HOTAIR（si-HOTAIR）序列的siRNA在人胶质瘤中的表达。进一步评价HOTAIR表达下调对人GSC增殖、侵袭和体内致瘤性的影响及其分子机制。MTT法和流式细胞仪分析结果显示HOTAIR表达下调可抑制细胞增殖并诱导细胞周期阻滞。Transwell实验表明HOTAIR表达下调导致GSC侵袭能力降低。此外，磁性纳米颗粒介导的HOTAIR的低表达有效地降低了胶质瘤干细胞在体内的致瘤能力。此外，qRT-PCR和western blot分析的结果表明，HOTAIR表达下调显著增加GSC中PDCD 4的表达，除了降低CCND 1和CDK 4的表达之外。深入的机制分析表明，HOTAIR表达的下调减少了下游分子EZH 2和LSD 1的募集，从而在转录水平上激活了PDCD 4的表达。总之，HOTAIR表达下调可有效促进PDCD 4的表达，从而抑制人GSC的增殖、侵袭和体内致瘤性。
Glioma stem cells (GSCs) are a special subpopulation of glioma cells that are key to the sensitivity of tumors to treatments and to the possibility of tumor recurrence. Identifying new strategies that inhibit the growth of GSCs are therefore important for developing novel therapies for glioblastoma multiforme (GBM). In this study, CD133+ human glioma stem cells were isolated and cultured. Magnetic nanoparticles were used to mediate the expression of siRNAs targeting the HOTAIR (si-HOTAIR) sequence in human gliomas. Effect of downregulation of HOTAIR expression on proliferation, invasion and in vivo tumorigenicity of human GSCs and underlying molecular mechanisms were further evaluated. The results of the MTT assay and flow cytometric analysis showed that downregulation of HOTAIR expression inhibited cell proliferation and induced cell cycle arrest. Transwell assays demonstrated that downregulation of HOTAIR expression resulted in a decrease in the invasive capability of GSCs. Moreover, magnetic nanoparticle-mediated low expression of HOTAIR effectively reduced the tumorigenic capacity of glioma stem cells in vivo. In addition, the results of qRT-PCR and western blot analysis demonstrated that downregulation of HOTAIR expression significantly increased the expression of PDCD4 in GSCs, in addition to reducing the expression of CCND1 and CDK4. An in-depth mechanistic analysis showed that downregulation of HOTAIR expression reduced the recruitment of downstream molecules, EZH2 and LSD1, thereby activating the expression of PDCD4 at the transcriptional level. In conclusion, downregulation of HOTAIR expression effectively promoted the expression of PDCD4, thereby inhibiting the proliferation, invasion and in vivo tumorigenicity of human GSCs.