目的 探讨长链非编码RNA H19(LncRNA H19)在他莫昔芬治疗失败的乳腺癌细胞中的表达情况,并分析其对细胞侵袭能力的影响。方法 通过荧光定量PCR检测LncRNA H19在亲本细胞MCF-7W和他莫昔芬耐药细胞MCF-7R的表达水平,进一步使用siRNA干扰技术探讨LncRNA H19的生物学功能。通过转染siRNA H19来下调LncRNA H19的表达,以空载体和阴性对照组为对照组,荧光定量PCR检测转染效率和转染前后表皮间质转化相关转录因子Snail-2的表达水平变化。用Transwell法评估下调LncRNA H19表达后对MCF-7R细胞侵袭能力的影响。结果 与MCF-7W细胞相比,MCF-7R细胞中H19lncRNA呈现高水平表达状态。且MCF-7R中EMT相关基因Snail2的表达均明显高于MCF-7W细胞株。siRNA H19转染MCF-7R细胞24 h后, LncRNA H19和Snail 2 mRNA表达水平均明显下调(P<0.01)。Transwell实验显示,通过转染siRNA H19下调LncRNA H19表达可抑制细胞的侵袭能力(P<0.01)。结论 LncRNA H19在他莫昔芬耐药乳腺癌细胞中高表达,而下调其表达可减弱耐他莫昔芬乳腺癌细胞的侵袭能力,H19成为未来治疗耐他莫昔芬乳腺癌转移的药物靶点。
Abstract
OBJECTIVE To investigate the expression profile of long non-coding RNA H19 (LncRNA H19) in tamoxifen-resistant breast cancer cell lines, and to study its biological functions on cell invasion. METHODS Quantitative reverse-transcription PCR(Q-PCR) was performed to detect the expression of LncRNA H19 in MCF-7W and MCF-7R cells. To further explore its biological function, RNA interference was applied. siRNA H19 was transfected to down-regulate H19 expression, and transfection efficiency was evaluated by Q-PCR. The epidermal mesenchymal transition-related transcription gene Snail 2 was also evaluated by Q-PCR. Transwell assay was performed to evaluate the effect of H19 expression on invasion potential of MCF-7R cells. RESULTS Compared with MCF-7W cells, MCF-7R cells exhibit a relatively high expression of lncRNA H19 and Snail2. LncRNA H19 expression was down-regulated in MCF-7R after transfection of siRNA H19 for 24 h. Snail 2 mRNA expression was significantly inhibited after down regulating H19 expression (P<0.01). Transwell assay indicated that inhibition of LncRNA H19 by siRNA H19 could repress cell invasion (P<0.01). CONCLUSION The expression of LncRNA H19 is significantly up-regulated in MCF-7R, and its downregulation attenuated the invasion behavior and related gene expression of tamoxifen-resistant breast cancer cell. H19 may be a drug treatment target of tamoxifen-resistant breast cancer metastasis.
关键词
长链非编码RNA H19 /
侵袭 /
表皮间质转化
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Key words
long noncoding RNA H19 /
invasion /
epidermal-mesenchymal transition
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中图分类号：
R965
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参考文献
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脚注
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基金
国家自然科学基金资助项目(81403021);福建省卫生与计划生育委员会医学创新课题资助项目(2014-CX-5);福建省立医院优秀青年培养项目(2014YNQN02)
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