HOST2 lncRNA靶向结合miR-211干预卵巢癌细胞转移的机制研究  

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

作　　者：热孜亚·库尔班 古力米热·布然江[1] 古扎丽努尔·阿不力孜[1] REZIYA·Kuerban;GULIMIRE·Buranjiang;GUZALINUER·Abulizi(Department of Gynaecology, affiliated Tumor Hospital of Xinjiang Medical University, Xinjiang, Urumqi 830011, China)

机构地区：[1]新疆医科大学附属肿瘤医院妇科,新疆乌鲁木齐830011

出　　处：《河北医科大学学报》2020年第9期1058-1064,共7页Journal of Hebei Medical University

基　　金：新疆维吾尔自治区自然科学基金项目(2015211C128)。

摘　　要：目的探讨HOST2 lncRNA靶向结合miR-211干预卵巢癌细胞转移的机制。方法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。结果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)。结论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。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.

关 键 词：卵巢肿瘤 HOST2 lncRNA miR-211 

分 类 号：R737.31[医药卫生—肿瘤]