HOST2 lncRNA靶向结合miR-211干预卵巢癌细胞转移的机制研究

目的〖KG*2〗探讨HOST2 lncRNA靶向结合miR-211干预卵巢癌细胞转移的机制。〖HTH〗方法〖KG*2〗实时荧光定量聚合酶链反应（quantitative real-time polymerase chain reaction，qRT-PCR）检测7种卵巢癌细胞系中HOST2 lncRNA及miR-211表达水平，选取其中差异表达最高卵巢癌细胞作为候选细胞系；双荧光素酶报告基因实验检测卵巢癌细胞中HOST2 lncRNA与miR-211间靶向相关性。将细胞分为转染无序HOST2 lncRNA及miR-211对照物的对照组、转染HOST2 lncRNA过表达载体的HOST2组、共转染HOST2 lncRNA与miR-211 mimic的（HOST2+miR-211）组。细胞划痕实验、Transwell小室实验及裸鼠腹腔移植瘤实验分别检测HOST2 lncRNA靶向miR-211对卵巢癌细胞体内外转移能力的影响；Western Blot检测卵巢癌细胞内转移相关蛋白的表达变化。〖HTH〗结果〖KG*2〗HOST2 lncRNA在7种卵巢癌细胞中显著高表达，miR-211在7种卵巢癌细胞中显著低表达，二者表达存在显著负相关关系，其中SKOV3细胞中HOST2 lncRNA及miR-211表达差异最高（P＜0.01）。双荧光素酶报告实验显示野生型HOST2报告基因质粒共转染miR-211 mimic后SKOV3细胞内荧光素酶活性较对照组受到显著抑制；过表达HOST2 lncRNA促进细胞划痕愈合、体外侵袭和迁移及裸鼠体内腹腔移植瘤的转移，反之共转染miR-211 mimic后抑制过表达HOST2 lncRNA促进的细胞体内外的转移；Western Blot结果显示过表达HOST2 lncRNA显著提高细胞内SOX4、Snail1及N-cadherin蛋白表达，下调E-cadherin蛋白表达，反之共转染miR-211 mimic后逆转上述蛋白表达变化（P＜0.01）。〖HTH〗结论〖KG*2〗HOST2 lncRNA可能通过作为内源竞争RNA竞争性结合miR-211促进卵巢癌细胞体内外的转移。

Mechanism of HOST2 lncRNA targeting miR-211 in ovarian cancer cell metastasis

ObjectiveTo investigate the mechanism of HOST2 lncRNA targeting miR-211 in ovarian cancer cell metastasis. MethodsQuantitative real-time polymerase chain reaction(qRT-PCR) was used to detect the expression of HOST2 lncRNA and miR-211 in seven kinds of ovarian cancer cells. Dual luciferase reporter gene assay was used to detecte the targeted correlation between of miR-211 and HOST2 lncRNA. The candidate cells were divided into 3 groups: the negative control group, the HOST2 lncRNA overexpression group and both HOST2 lncRNA and miR-211 overexpression group. Cell scratch assay, transwell invasion and migration assays and nude mice xenograft tumor assays were conducted to assess the effects of HOST2 lncRNA and miR-211 on cell metastasis capabilities in vivo and in vitro after transfection. Western Blot was used to detect the expression changes of metastasis-associated proteins in ovarian cancer cells. ResultsHOST2 lncRNA was highly expressed in seven ovarian cancer cells while miR-211 was significantly down-regulated in these cells. And there was a significant negative correlation between HOST2 lncRNA and miR-211. Dual luciferase reporter assay showed that the luciferase activity of SKOV3 cells was significantly inhibited in the SKOV3 cells after co-transfection of the HOST2-WT with miR-211 mimic. Overexpression of HOST2 lncRNA promoted cell scratch healing, invasion and migration in vitro and metastasis of peritoneal xenografts in vivo. In turn, co-transfection of miR-211 mimic reversed this phenomenon. Western Blot results showed that overexpression of HOST2 lncRNA significantly increased the expression of SOX4, Snail1 and N-cadherin proteins and decreased the expression of E-cadherin protein, whereas co-transfection of miR-211 mimic reversed the expression of the proteins above. ConclusionHOST2 lncRNA may promote the metastasis of ovarian cancer cells in vivo and in vitro by competitively binding to miR-211 as competing endogenous RNAs.