上海口腔医学

上海口腔医学 ›› 2023, Vol. 32 ›› Issue (1): 85-90.doi: 10.19439/j.sjos.2023.01.016

• 论著 • 上一篇    下一篇

LncRNA RUNX1-IT1对恶性多形性腺瘤miR-195/CyclinD1的调控作用探讨

魏军水, 孙鑫, 徐金标, 陈逸达   

  1. 台州市第一人民医院 口腔科,浙江 台州 318020
  • 收稿日期:2022-07-16 修回日期:2022-10-17 出版日期:2023-02-25 发布日期:2023-06-12
  • 通讯作者: 魏军水,E-mail: weijunshui@163.com
  • 作者简介:魏军水（1980-）,男,硕士,副主任医师
  • 基金资助:
    台州市科技计划项目（1901ky61）

LncRNA RUNX1-IT1 regulating malignant pleomorphic adenoma via mir-195/CyclinD1

WEI Jun-shui, SUN Xin, XU Jin-biao, CHEN Yi-da   

  1. Department of Stomatology, Taizhou First People's Hospital. Taizhou 318020, Zhejiang Province, China
  • Received:2022-07-16 Revised:2022-10-17 Online:2023-02-25 Published:2023-06-12

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摘要： 目的: 探讨恶性多形性腺瘤(malignant pleomorphic adenoma, MPA)细胞中长链非编码RNA(long non-coding RNA, LncRNA)RUNX1-IT1对微小RNA-195(microRNA-195, miR-195) /细胞周期蛋白D1(CyclinD1)的调控作用。方法: 收集手术切除的MPA组织及癌旁组织,检测LncRNA RUNX1-IT1、miR-195、CyclinD1 mRNA的表达水平,分析三者与MPA临床病理的关系。培养MPA细胞系SM-AP1,转染阴性对照(NC)siRNA、LncRNA RUNX1-IT1 siRNA、miR-NC、miR-195抑制物,检测细胞增殖水平A490及miR-195、CyclinD1的表达水平,分析LncRNA RUNX1-IT1靶向miR-195、miR-195靶向CyclinD1的关系。采用SPSS 21.0软件包对数据进行统计学分析。结果: MPA中LncRNA RUNX1-IT1、CyclinD1的表达水平显著高于癌旁组织,miR-195的表达水平显著低于癌旁组织(P<0.05)。LncRNA RUNX1-IT1与miR-195呈负相关、与CyclinD1呈正相关,miR-195与CyclinD1呈负相关。肿瘤直径≥3 cm、复发、远处转移的MPA癌组织中,LncRNA RUNX1-IT1、CyclinD1表达更高(P<0.05),miR-195表达更低(P<0.05)。敲低LncRNA RUNX1-IT1后,SM-AP1细胞的A490水平、CyclinD1表达水平降低,miR-195表达水平增加(P<0.05)。LncRNA RUNX1-IT1通过直接靶向作用于SM-AP1细胞中的miR-195,miR-195通过直接靶向作用于SM-AP1细胞中的CyclinD1。抑制miR-195后,敲低LncRNA RUNX1-IT1降低A490水平及CyclinD1表达水平的作用减弱（P<0.05）。结论: LncRNA RUNX1-IT1可能通过调控miR-195/CyclinD1表达,参与MPA的发生、发展。

关键词: 恶性多形性腺瘤, LncRNA RUNX1-IT1, miR-195, CyclinD1, 增殖, 靶基因

Abstract: PURPOSE: To investigate the regulation of long non-coding RNA (LncRNA) RUNX1-IT1 on microrna (mir-195)/CyclinD1 (CyclinD1) in malignant pleomorphic adenoma (MPA). METHODS: The MPA tissues and para-carcinoma tissues were collected and the expression levels of LncRNA RUNX1-IT1, miR-195 and CyclinD1 mRNA were detected, the correlation and clinical pathology of MPA was analyzed and compared. MPA cell line SM-AP1 was cultured and transfected with negative control(NC) siRNA, LncRNA RUNX1-IT1siRNA, miR-NC and miR-195 inhibitor. Cell proliferation level A490 and expression levels of miR-195 and CyclinD1 were detected. LncRNA RUNX1-IT1 targeting miR-195 and miR-195 targeting CyclinD1 were analyzed by dual luciferase reporter gene assay. SPSS 21.0 software package was used for data analysis. RESULTS: The expression level of LncRNA RUNX1-IT1 and CyclinD1 in MPA were higher than those in para tumor tissues, and the expression level of miR-195 was lower than that in para tumor tissues(P<0.05). LncRNA RUNX1-IT1was negatively correlated with miR-195, positively correlated with CyclinD1, and miR-195 was negatively correlated with CyclinD1. The expression of LncRNA RUNX1-IT1 and CyclinD1 in MPA tissue with tumor diameter≥3 cm, recurrence and distant metastasis increased(P<0.05), while the expression of miR-195 decreased(P<0.05). After knockdown of LncRNA RUNX1-IT1, A490 level and CyclinD1 expression level decreased, while miR-195 expression level increased(P<0.05). miR-195 decreased the fluorescence activity of LncRNA RUNX1-IT1 and CyclinD1 reporter genes(P<0.05). After miR-195 was inhibited, the effect of LncRNA RUNX1-IT1 knockdown on decreasing A490 level and CyclinD1 expression level weakened(P<0.05). CONCLUSIONS: LncRNA RUNx1-IT1 may participate in the development of MPA by regulating the expression of miR-195/CyclinD1.

Key words: Malignant pleomorphic adenoma, LncRNA RUNX1-IT1, miR-195, CyclinD1, Proliferation, Target gene

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