河北医科大学学报

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HOST2 lncRNA靶向结合miR-211干预卵巢癌细胞转移的机制研究

  

  1. 新疆医科大学附属肿瘤医院妇科,新疆 乌鲁木齐 830011
  • 出版日期:2020-09-25 发布日期:2020-09-27
  • 作者简介:热孜亚·库尔班(1982-),女,维吾尔族,新疆乌鲁木齐人,新疆医科大学附属肿瘤医院主治医师,医学博士研究生,从事妇科肿瘤疾病诊治研究。
  • 基金资助:
    新疆维吾尔自治区自然科学基金项目(2015211C128)

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

  1. Department of Gynaecology, affiliated Tumor Hospital of Xinjiang Medical University, Xinjiang, Urumqi 830011, China
  • Online:2020-09-25 Published:2020-09-27

摘要: 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。

关键词: 卵巢肿瘤, HOST2 lncRNA, miR-211

Abstract: Objective  To investigate the mechanism of HOST2 lncRNA targeting miR-211 in ovarian cancer cell metastasis.
  Methods  Quantitative 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.
  Results  HOST2 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.
  Conclusion  HOST2 lncRNA may promote the metastasis of ovarian cancer cells in vivo and in vitro by competitively binding to miR-211 as competing endogenous RNAs.

Key words: ovarian neoplasms;HOST2 lncRNA, miR-211