Nanoparticle Delivery of miR-34a Eradicates Long-term-cultured Breast Cancer Stem Cells via Targeting C22ORF28 Directly
Nanoparticle Delivery of miR-34a Eradicates Long-term-cultured Breast Cancer Stem Cells via Targeting C22ORF28 Directly
复制标题
miR-34a 的纳米颗粒递送通过直接靶向 C22ORF28 消除长期培养的乳腺癌干细胞
DOI：
10.7150/thno.20771
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发表时间：
2017-01-01
期刊：
影响因子：
12.4
通讯作者：
Xie, Xiaoming
中科院分区：
文献类型：
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作者：
Lin, Xiaoti;Chen, Weiyu;Xie, Xiaoming
基本原理：肿瘤干细胞（cancer stem cells，CSCs）具有自我更新能力、广泛的分化潜能和对常规治疗的抵抗力，被认为是肿瘤耐药和转移的种子。这是一种积极主动的癌症治疗策略，以消除CSC。研究方法：肿瘤组织来源的乳腺CSC（BCSC），包括XM 322和XM 607，通过荧光激活细胞分选（FACS）分离;而细胞系来源的BCSC，包括MDA-MB-231.SC和MCF-7.SC，通过磁激活细胞分选（MACS）纯化。在多个乳腺细胞系中进行了微小RNA和mRNA表达阵列谱分析。上述纳米颗粒按照标准分子克隆方案构建。组织芯片分析已被用于研究217例临床乳腺癌标本。结果：在这里，我们成功建立了四个长期维持的BCSC，保留了其引发肿瘤的生物学特性。我们的微阵列和qRT-PCR分析发现，miR-34 a是研究BCSC最明显的microRNA。我们建立了hTERT启动子驱动的VISA递送miR-34 a（TV-miR-34 a）质粒，其可以诱导BCSC中miR-34 a的高通量表达。TV-miR-34 a在体外可显著抑制长期培养的BCSC的致瘤特性，在体内可安全有效地抑制BCSC的增殖。TV-miR-34 a与多西他赛（侵袭性乳腺癌的标准疗法）协同作用，作为BCSC抑制剂。进一步的机制研究表明，TV-miR-34 a直接阻止C22 ORF 28积累，这消除了克隆形成和肿瘤生长，并与乳腺癌患者中的低miR-34和高C22 ORF 28水平相关。结论：综上所述，我们从临床标本或细胞系中获得了四个长期维持的BCSC，这将大大有利于乳腺癌患者的研究进展。我们进一步开发了非病毒TV-miR-34 a质粒，该质粒具有很大的潜力，可作为乳腺癌治疗的临床应用。
Rationale: Cancer stem cells (CSCs) have been implicated as the seeds of therapeutic resistance and metastasis, due to their unique abilities of self-renew, wide differentiation potentials and resistance to most conventional therapies. It is a proactive strategy for cancer therapy to eradicate CSCs. Methods: Tumor tissue-derived breast CSCs (BCSC), including XM322 and XM607, were isolated by fluorescence-activated cell sorting (FACS); while cell line-derived BCSC, including MDA-MB-231.SC and MCF-7.SC, were purified by magnetic-activated cell sorting (MACS). Analyses of microRNA and mRNA expression array profiles were performed in multiple breast cell lines. The mentioned nanoparticles were constructed following the standard molecular cloning protocol. Tissue microarray analysis has been used to study 217 cases of clinical breast cancer specimens. Results: Here, we have successfully established four long-term maintenance BCSC that retain their tumor-initiating biological properties. Our analyses of microarray and qRT-PCR explored that miR-34a is the most pronounced microRNA for investigation of BCSC. We establish hTERT promoter-driven VISA delivery of miR-34a (TV-miR-34a) plasmid that can induce high throughput of miR-34a expression in BCSC. TV-miR-34a significantly inhibited the tumor-initiating properties of long-term-cultured BCSC in vitro and reduced the proliferation of BCSC in vivo by an efficient and safe way. TV-miR-34a synergizes with docetaxel, a standard therapy for invasive breast cancer, to act as a BCSC inhibitor. Further mechanistic investigation indicates that TV-miR-34a directly prevents C22ORF28 accumulation, which abrogates clonogenicity and tumor growth and correlates with low miR-34 and high C22ORF28 levels in breast cancer patients. Conclusion: Taken together, we generated four long-term maintenance BCSC derived from either clinical specimens or cell lines, which would be greatly beneficial to the research progress in breast cancer patients. We further developed the non-viral TV-miR-34a plasmid, which has a great potential to be applied as a clinical application for breast cancer therapy.