国际口腔医学杂志

国际口腔医学杂志 ›› 2024, Vol. 51 ›› Issue (1): 60-67.doi: 10.7518/gjkq.2024007

• 论著 • 上一篇    下一篇

环状RNA hsa_circ_0085576调控微小RNA-498/B细胞特异性莫洛尼鼠白血病病毒整合位点1轴对口腔鳞状细胞癌细胞迁移和侵袭的影响

李立恒1(),王蕊1,王晓明1,张智轶1,张璇1,安峰1,王芹1,张凡2   

  1. 1.河北北方学院附属第一医院口腔科 张家口 075000
    2.河北北方学院附属第一医院病理科 张家口 075000
  • 收稿日期:2023-06-15 修回日期:2023-09-26 出版日期:2024-01-01 发布日期:2024-01-10
  • 通讯作者: 李立恒
  • 作者简介:李立恒，主治医师，硕士，Email：jugrmu@163.com
  • 基金资助:
    2021年度河北省医学科学研究课题计划(20210802)

Effects of circular RNA hsa_circ_0085576 on cell migration and invasion of oral squamous cell carcinoma by regulating the microRNA-498/B-cell-specific Moloney murine leukemia virus integration site 1 axis

Li Liheng1(),Wang Rui1,Wang Xiaoming1,Zhang Zhiyi1,Zhang Xuan1,An Feng1,Wang Qin1,Zhang Fan2   

  1. 1.Dept. of Stomatology, the First Affiliated Hospital of Hebei North University, Zhangjiakou 075000, China
    2.Dept. of Pathology, the First Affiliated Hospital of Hebei North University, Zhangjiakou 075000, China
  • Received:2023-06-15 Revised:2023-09-26 Online:2024-01-01 Published:2024-01-10
  • Contact: Liheng Li
  • Supported by:
    Hebei Province 2021 Annual Medical Science Research Project(20210802)

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摘要：

目的 探究环状RNA hsa_circ_0085576对口腔鳞状细胞癌（OSCC）细胞迁移和侵袭的影响及其分子机制。 方法 使用实时荧光定量聚合酶链反应（qRT-PCR）和蛋白印迹法检测OSCC细胞中hsa_circ_0085576、微小RNA-498（miR-498）以及B细胞特异性莫洛尼鼠白血病病毒整合位点1（BMI-1）的表达水平。使用CCK-8、划痕实验、Transwell实验以及qRT-PCR、蛋白印迹法分别检测SCC-15细胞增殖活力、迁移及侵袭能力以及相关基因和蛋白的相对表达量。 结果 OSCC细胞中hsa_circ_0085576与BMI-1表达上调，miR-498表达下调（P<0.05）。下调hsa_circ_0085576表达或过表达miR-498后SCC-15细胞的增殖活性、划痕愈合率、侵袭细胞数目以及细胞周期蛋白D1、波形蛋白表达水平下调，miR-498和E-钙黏蛋白表达水平上调（P<0.05）；抑制miR-498表达可减弱下调hsa_circ_0085576表达对OSCC细胞增殖、迁移及侵袭的抑制作用；上调BMI-1表达可减弱过表达miR-498对OSCC细胞增殖、迁移及侵袭的抑制作用。 结论 下调hsa_circ_0085576表达可通过激活miR-498/BMI-1轴抑制OSCC细胞增殖、迁移及侵袭。

关键词: 环状RNA hsa_circ_0085576, 微小RNA-498, B细胞特异性莫洛尼鼠白血病病毒整合位点1, 口腔鳞状细胞癌

Abstract:

Objective This study aims to explore the effects of circular RNA hsa_circ_0085576 on cell migration and invasion of oral squamous cell carcinoma (OSCC) and the underlying molecular mechanism. Methods Circular RNA hsa_circ_0085576, microRNA-498 (miR-498), and B-cell-specific Moloney murine leukemia virus integration site 1 (BMI-1) in the cells of OSCC were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analyses. CCK-8, scratch test, Transwell test, qRT-PCR, and Western blot were used to detect the proliferation, migration, and invasion ability of SCC-15 cells and the expression of related genes and proteins. Results The expression of hsa_circ_0085576 and BMI-1 in the cells of OSCC was upregulated, and that of miR-498 was downregulated (P<0.05). The proliferative activity, scratch healing rate, number of invasive cells, and expression levels of cyclinD1 and vimentin proteins of SCC-15 cells were downregulated, whereas the expression levels of miR-498 and E-cadherin proteins were upregulated (P<0.05). Inhibition of miR-498 expression weakened the inhibitory effect of downregulating hsa_circ_0085576 on the proliferation, migration, and invasion of OSCC cells. Upregulation of BMI-1 expression attenuated the inhibitory effect of overexpressed miR-498 on the proliferation, migration, and invasion of OSCC cells. Conclusion Downregulation of hsa_circ_0085576 expression inhibited the proliferation, migration, and invasion of OSCC cells by activating the miR-498/BMI-1 axis.

Key words: circular RNA hsa_circ_0085576, microRNA-498, B-cell-specific Moloney murine leukemia virus integration site 1, oral squamous cell carcinoma

中图分类号: 

  • R730.2

表 1

qRT-PCR引物序列"

基因上游引物（5’—3’）下游引物（5’—3’）
hsa_circ_0085576CTTTGCAGTGAGCCATGGGAGAGTCGTCTCTCCTGTCACG
BMI-1CTGGTTGCCCATTGACAGCCAGAAAATGAATGCGAGCCA
GAPDHCTCTGCTCCTCCTGTTCGACGCGCCCAATACGACCAAATC
miR-498GGTTTGAAGCCAGGCGGTTTCCAGTGCAGGGTCCGAGGTAT
U6GTGCTCGCTTCGGCAGCACATATACAAAAATATGGAACGCTTCACGAATTTG

图1

不同OSCC细胞中hsa_circ_0085576、miR-498和BMI-1表达水平A：hsa_circ_0085576相对表达量；B：miR-498相对表达量；C：BMI-1 mRNA相对表达量；D、E：Western blot测定BMI-1蛋白相对表达量。*：与NOK组相比，P<0.05。"

图2

下调hsa_circ_0085576表达对BMI-1蛋白表达的影响"

表 2

下调hsa_circ_0085576表达对miR-498与BMI-1表达的影响"

组别hsa_circ_0085576miR-498BMI-1蛋白
si-NC1.02±0.071.00±0.061.29±0.18
干扰10.52±0.04*1.55±0.11*0.57±0.06*
干扰20.24±0.00*2.15±0.12*0.46±0.04*
干扰30.31±0.02*1.96±0.14*0.53±0.03*

图3

下调hsa_circ_0085576和miR-498表达对SCC-15细胞增殖、迁移、侵袭的影响A：CCK-8检测细胞增殖活力；B、C：划痕实验测定细胞迁移；D、E：Tranwell实验测定细胞侵袭；F：qRT-PCR检测miR-498相对表达水平；G、H：Western blot测定CyclinD1、Vimentin、E-cadherin以及BMI-1的蛋白表达；组别：1为si-NC组，2为干扰2组，3为干扰2+miR inhibitor组，4为干扰2+miR-498 inhibitor组。*：与si-NC组相比，P<0.05；#：与干扰2+miR inhibitor组相比，P<0.05。"

图4

过表达miR-498和BMI-1对SCC-15细胞增殖、迁移、侵袭的影响A：CCK-8检测细胞增殖活力；B、C：划痕实验测定细胞迁移；D、E：Tranwell实验测定细胞侵袭；F、G：Western blot测定BMI-1、CyclinD1、Vimentin以及E-cadherin的蛋白表达；组别：1为miR mimics组，2为miR-498 mimics组，3为miR-498 mimics+Ctrl组，4为miR-498 mimics+BMI-1组。与miR mimics组相比，*P<0.05；与miR-498 mimics+Ctrl组相比，#P<0.05。"

图5

hsa_circ_0085576、miR-498和BMI-1的靶向关系A：预测的hsa_circ_0085576与miR-498的结合位点，红色表示突变的结合位点；B：hsa_circ_0085576与miR-498的双荧光素酶检测结果；C：预测的miR-498与BMI-1的结合位点，红色表示突变的结合位点；D：miR-498与BMI-1的双荧光素酶检测结果。与miR-NC组相比，*P<0.05。"

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