Department of Radiation Medicine, Fourth Military Medical University, 169 Changle Western Road, Xi'an, Shaanxi 710032, China.
State Key Laboratory of Cancer Biology, Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi'an, Shaanxi 710032, China.
Cancer Cell Int. 2013 Nov 27;13(1):118. doi: 10.1186/1475-2867-13-118.
The mechanisms responsible for cervical cancer radioresistance are still largely unexplored. The present study aimed to identify miRNAs associated with radioresistance of cervical cancer cells.
The radioresistant cervical cancer cell variants were established by repeated selection with irradiation. The miRNA profiles of radioresistant cells and their corresponding controls were analyzed and compared using microarray. Differentially expressed miRNAs were confirmed by quantitative real-time PCR. Cervical cancer cells were transfected with miRNA-specific mimics or inhibitors. Radiosensitivity of cervical cancer cells were determined using colony-forming assay.
Among the differentially expressed miRNAs, 20 miRNAs showed the similar pattern of alteration (14 miRNAs were overexpressed whilst 6 were suppressed) in all three radioresistant cervical cancer cell variants compared to their controls. A miRNA signature consisting of 4 miRNAs (miR-630, miR-1246, miR-1290 and miR-3138) exhibited more than 5 folds of increase in radioresistant cells. Subsequent analysis revealed that these four miRNAs could be up-regulated in cervical cancer cells by radiation treatment in both time-dependent and dose-dependent manners. Ectopic expression of each of these 4 miRNAs can dramatically increase the survival fraction of irradiated cervical cancer cells. Moreover, inhibition of miR-630, one miRNA of the specific signature, could reverse radioresistance of cervical cancer cells.
The present study indicated that miRNA is involved in radioresistance of human cervical cancer cells and that a specific miRNA signature consisting of miR-630, miR-1246, miR-1290 and miR-3138 could promote radioresistance of cervical cancer cells.
导致宫颈癌放射抵抗的机制在很大程度上仍未得到探索。本研究旨在鉴定与宫颈癌细胞放射抵抗相关的 miRNA。
通过反复照射选择建立耐辐射的宫颈癌细胞株。采用微阵列分析比较耐辐射细胞及其相应对照的 miRNA 谱。通过实时定量 PCR 验证差异表达的 miRNA。用 miRNA 特异性模拟物或抑制剂转染宫颈癌细胞。采用集落形成实验测定宫颈癌细胞的放射敏感性。
在差异表达的 miRNA 中，有 20 个 miRNA 在所有三种耐辐射的宫颈癌细胞株中与对照相比呈现相似的改变模式（14 个 miRNA 过表达，而 6 个 miRNA 受抑制）。一个由 4 个 miRNA（miR-630、miR-1246、miR-1290 和 miR-3138）组成的 miRNA 特征在耐辐射细胞中表达增加了 5 倍以上。随后的分析表明，这些 miRNA 可以通过辐射处理在时间和剂量依赖性方式在宫颈癌细胞中上调。这些 miRNA 中的每一个的异位表达都可以显著增加照射的宫颈癌细胞的存活分数。此外，miR-630 的抑制，即特异性特征之一，可以逆转宫颈癌细胞的放射抵抗性。
本研究表明 miRNA 参与了人宫颈癌细胞的放射抵抗，由 miR-630、miR-1246、miR-1290 和 miR-3138 组成的特异性 miRNA 特征可促进宫颈癌细胞的放射抵抗。
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