MiRNA Transcriptome Profiling of Spheroid-Enriched Cells with Cancer Stem Cell Properties in Human Breast MCF-7 Cell Line.
MiRNA Transcriptome Profiling of Spheroid-Enriched Cells with Cancer Stem Cell Properties in Human Breast MCF-7 Cell Line.
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DOI：
10.7150/ijbs.12777
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发表时间：
2016
影响因子：
9.2
通讯作者：
Cheong SK
中科院分区：
文献类型：
--
作者：
Boo L;Ho WY;Ali NM;Yeap SK;Ky H;Chan KG;Yin WF;Satharasinghe DA;Liew WC;Tan SW;Ong HK;Cheong SK
乳腺癌是全球癌症相关死亡的第二大原因，因为大多数患者经常遭受癌症复发。其原因通常归因于癌症干细胞（CSC）的存在。近年来的研究发现，microRNA（miRNA）的表达异常与乳腺癌的复发和转移密切相关。然而，没有具体的研究已经全面表征了富含球状体的乳腺细胞中的CSC特征和miRNA转录组。本研究描述了无血清条件下球形MCF-7细胞的产生及其CSC特性的综合表征。随后，使用下一代测序（NGS）评估球状体富集的CSC细胞与其亲本细胞之间的miRNA表达差异。我们的研究结果表明，MCF-7球体细胞富含CSCs特性，表现为自我更新能力、CSCs标志物表达增加以及对化疗药物的耐药性增加。此外，球状体富集的CSC具有更强的细胞增殖、迁移、侵袭和伤口愈合能力。使用miRNA-NGS在球状体和亲本细胞之间鉴定了总共134个显著（p<0.05）差异表达的miRNA。MiRNA-NGS分析显示25个上调和109个下调的miRNAs，其中包括一些先前报道的乳腺CSC调控中的miRNAs。一些以前未在乳腺癌中报道的miRNA（miR-4492、miR-4532、miR-381、miR-4508、miR-4448、miR-1296和miR-365 a）被发现与乳腺癌化学抗性和自我更新能力显示出潜在的关联。基因本体（GO）分析表明，预测的基因丰富的代谢过程的调控，基因表达，DNA结合，激素受体结合。从GO结果推断出的相应途径分析与乳腺癌中信号通路的功能、自我更新能力、化疗耐药性、肿瘤发生、细胞骨架蛋白和转移密切相关。基于这些结果，我们提出本研究中鉴定的某些miRNAs可作为新的潜在生物标志物用于乳腺癌干细胞诊断和靶向治疗。
Breast cancer is the second leading cause of cancer-related mortality worldwide as most patients often suffer cancer relapse. The reason is often attributed to the presence of cancer stem cells (CSCs). Recent studies revealed that dysregulation of microRNA (miRNA) are closely linked to breast cancer recurrence and metastasis. However, no specific study has comprehensively characterised the CSC characteristic and miRNA transcriptome in spheroid-enriched breast cells. This study described the generation of spheroid MCF-7 cell in serum-free condition and the comprehensive characterisation for their CSC properties. Subsequently, miRNA expression differences between the spheroid-enriched CSC cells and their parental cells were evaluated using next generation sequencing (NGS). Our results showed that the MCF-7 spheroid cells were enriched with CSCs properties, indicated by the ability to self-renew, increased expression of CSCs markers, and increased resistance to chemotherapeutic drugs. Additionally, spheroid-enriched CSCs possessed greater cell proliferation, migration, invasion, and wound healing ability. A total of 134 significantly (p<0.05) differentially expressed miRNAs were identified between spheroids and parental cells using miRNA-NGS. MiRNA-NGS analysis revealed 25 up-regulated and 109 down-regulated miRNAs which includes some miRNAs previously reported in the regulation of breast CSCs. A number of miRNAs (miR-4492, miR-4532, miR-381, miR-4508, miR-4448, miR-1296, and miR-365a) which have not been previously reported in breast cancer were found to show potential association with breast cancer chemoresistance and self-renewal capability. The gene ontology (GO) analysis showed that the predicted genes were enriched in the regulation of metabolic processes, gene expression, DNA binding, and hormone receptor binding. The corresponding pathway analyses inferred from the GO results were closely related to the function of signalling pathway, self-renewability, chemoresistance, tumorigenesis, cytoskeletal proteins, and metastasis in breast cancer. Based on these results, we proposed that certain miRNAs identified in this study could be used as new potential biomarkers for breast cancer stem cell diagnosis and targeted therapy.