Sun Ning-xia, Ye Chen, Zhao Qian, Zhang Qing, Xu Chen, Wang Shao-bing, Jin Zhi-jun, Sun Shu-han, Wang Fang, Li Wen
Department of Obstetrics and Gynaecology, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai, China.
Department of Medical Genetics, Second Military Medical University, Shanghai, China.
PLoS One. 2014 Jul 9;9(7):e100340. doi: 10.1371/journal.pone.0100340. eCollection 2014.
In recent years, long noncoding RNAs (lncRNAs) have been demonstrated to play key roles in tumorgenesis. However, the contributions of lncRNAs to cervical cancer (CC) remain largely unknown. In this study, differentially expressed lncRNAs and mRNAs in cervical cancer and paired peritumoral tissues were detected by transcriptome microarray analysis. We found 708 probe sets of lncRNAs increased and 836 probe sets decreased in CC tissues, while 1288 mRNA differential probe sets increased and 901 mRNA probe sets decreased. The results were validated by quantitative real-time polymerase chain reaction (qPCR). Then, we found a specific differentially expressed lncRNA can physically bind to enhancer of zeste homolog2 (EZH2) by using RNA immunoprecipitation. We termed it as EZH2-binding lncRNA in cervical cancer [lncRNA-EBIC]. Wound healing assays and Matrigel invasion assays were used to determine the function of this lncRNA by silencing it. We observed that the migration and invasion of cervical cancer cells in vitro were inhibited upon suppression of lncRNA-EBIC by siRNA. We also found that the association between lncRNA-EBIC and EZH2 was required for the repression of E-cadherin, which was a key molecular in the metastasis of cervical cancer. Conclusion: These results demonstrated that lncRNA-EBIC was an oncogenic lncRNA, which could promote tumor cell invasion in CC by binding to EZH2 and inhibiting E-cadherin expression.
近年来，长链非编码RNA（lncRNAs）已被证明在肿瘤发生中起关键作用。然而，lncRNAs对宫颈癌（CC）的作用仍 largely未知。在本研究中，通过转录组微阵列分析检测了宫颈癌组织和配对的肿瘤旁组织中差异表达的lncRNAs和mRNAs。我们发现CC组织中有708个lncRNAs探针集增加，836个探针集减少，而1288个mRNA差异探针集增加，901个mRNA探针集减少。结果通过定量实时聚合酶链反应（qPCR）进行验证。然后，我们发现一种特异性差异表达的lncRNA可以通过RNA免疫沉淀与zeste同源物2（EZH2）的增强子物理结合。我们将其命名为宫颈癌中与EZH2结合的lncRNA[lncRNA-EBIC]。通过沉默该lncRNA，使用伤口愈合试验和基质胶侵袭试验来确定其功能。我们观察到，用小干扰RNA（siRNA）抑制lncRNA-EBIC后，体外宫颈癌细胞的迁移和侵袭受到抑制。我们还发现lncRNA-EBIC与EZH2之间的关联是抑制E-钙黏蛋白所必需的，E-钙黏蛋白是宫颈癌转移中的关键分子。结论：这些结果表明lncRNA-EBIC是一种致癌性lncRNA，它可以通过与EZH2结合并抑制E-钙黏蛋白表达来促进CC中的肿瘤细胞侵袭。
