长链非编码RNA H19调控激素依赖型乳腺癌干细胞自我更新的作用及机制研究

Therapeutic effects and mechanistic regulation of H19 controlling of estrogen induction of breast cancer stem-like cells’ renewal

目的:探究长链非编码RNA H19在控制乳腺癌干细胞(breast cancer stem-like cells,BrCSC)分裂中的作用及机制。方法:H19临床前研究数据样本资料通过TCGA数据库分析实现。BrCSC自我更新能力通过微球体成球验证。功能性通路验证通过逆转录-聚合酶链反应、免疫印迹、荧光素酶报告基因实验等。结果:H19通过调节抑制Let-7c的功能发挥,进而激活BrCSC中雌激素维持的Wnt通路活性。Wnt通路激活进一步刺激H19增加,降低Let-7c的生物利用度,形成反馈环路。同理,恢复Let-7c的表达将减弱雌激素依赖的Wnt通路功能,进而抑制H19,抑制BrCSC的对称分裂。结论:H19通过与Let-7c结合并依赖的方式调控Wnt通路活性,形成H19/Let-7/Wnt反馈环路,在BrCSC的自我更新能力调控中起关键作用。

Objective:To explore the role and mechanism of long non-coding RNA H19 in controlling breast cancer stem-like cells(BrCSC)division.Methods:Sample data of H19 pre-clinical study were analyzed by TCGA database.The self-renewal ability was assessed by spheres formation.The mechanistic procedures were tested by RT-PCR,Western blot and Luciferase assay.Results:H19 activated the estrogen-maintained Wnt pathway activity in BrCSC by inhibiting the function of Let-7c.The activation of Wnt pathway further stimulated the increase of H19,reduced the bioavailability of Let-7c and formd a feedback loop.Similarly,restoring Let-7c expression would weaken the estrogen-dependent Wnt pathway,thereby inhibiting H19 and symmetrical division of BrCSC.Conclusion:H19 regulates Wnt pathway activity by binding and depending on Let-7c,and forms H19/Let-7/Wnt feedback loop,which plays a key role in the regulation of BrCSC's self-renewal ability.