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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。
Objective To investigate the expression of long-chain non-coding RNA H19 (LncRNA H19) in breast cancer cells that failed to be treated with tamoxifen and analyze its effect on cell invasiveness. Methods The expression of LncRNA H19 in parental MCF-7W and tamoxifen-resistant cells MCF-7R was detected by real-time PCR. The biological function of LncRNA H19 was further explored by siRNA interference. The expression of LncRNA H19 was down-regulated by transfection of siRNA H19, and the empty vector and negative control group were used as control group. The transfection efficiency and the expression level of Snail-2, a transcription factor involved in epidermal stromal transformation, were detected by real-time PCR. Transwell assay was used to evaluate the effect of down-regulating the expression of LncRNA H19 on the invasiveness of MCF-7R cells. Results Compared with MCF-7W cells, H19lncRNA showed a high level of expression in MCF-7R cells. The expression of EMT-related gene Snail2 in MCF-7R was significantly higher than that in MCF-7W cell line. The expression of LncRNA H19 and Snail 2 mRNA were down-regulated in siRNA-H19 transfected MCF-7R cells for 24 h (P <0.01). Transwell experiments showed that transfection of siRNA H19 down-regulated LncRNA H19 expression can inhibit cell invasion (P <0.01). Conclusions LncRNA H19 is overexpressed in tamoxifen-resistant breast cancer cells, and downregulation of LncRNA H19 may reduce the invasion ability of the tamoxifen-derived breast cancer cells. H19 is the target of future drug therapy for the metastasis of tamoxifen to breast cancer .