miR-219-5p靶向SOX5在口腔癌中的作用初探

周婷; 何影; 董晓函; 习杨彦彬; 陈波; 佟钧; 毛瑞

doi:10.12259/j.issn.2095-610X.S20230201

miR-219-5p靶向SOX5在口腔癌中的作用初探

doi: 10.12259/j.issn.2095-610X.S20230201

周婷¹,
何影^{1, 2},
董晓函³,
习杨彦彬³,
陈波⁴,
佟钧⁵,
毛瑞^1, ,

1.
昆明医科大学口腔医学院，云南昆明　650500
2.
武警云南省总队医院门诊部，云南昆明　650100
3.
昆明医科大学基础医学院，云南昆明　650500
4.
昆明医科大学科技成果孵化中心，云南昆明　650500
5.
昆明医科大学体育部，云南昆明　650500

基金项目: 云南省教育厅科学研究基金项目（2019J1292）

详细信息

作者简介:
周婷（1981～），女，云南楚雄人，医学博士，副教授，副主任医师，主要从事口腔正畸的临床诊疗与教学科研工作

何影与周婷对本文有同等贡献

通讯作者:
毛瑞，E-mail：1139714630@qq.com

中图分类号: R739.8
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出版历程
- 收稿日期: 2022-12-08
- 网络出版日期: 2023-01-31
- 刊出日期: 2023-02-25

Possible Roles of MiR-219-5p Targeting SOX5 in the Development of Oral Cancer

Ting ZHOU¹,
Ying HE^{1, 2},
Xiaohan DONG³,
Yanbin XIYANG³,
Bo CHEN⁴,
Jun TONG⁵,
Rui MAO^{1
, ,}

1.
School of Stomatology，Kunming Medical University，Kunming Yunnan 650500
2.
Outpatient Department，Yunnan Province Corps Hospital，Chinese People’s Armed Police Forces，Kunming Yunnan 650100
3.
Institute of Neuroscience，Basic Medical College
4.
Science and Technology Achievement Incubation Center
5.
Physical Education Department，Kunming Medical University，Kunming Yunnan 650500，China

摘要

摘要: 目的研究miR-219-5p 与SOX5在口腔鳞状细胞癌中是否存在差异性表达，并探讨miR-219-5p 对SOX5的靶向调控作用，方法运用口腔鳞状细胞癌细胞株SCC4及SCC9，人正常皮肤成纤维细胞系（HF）作为对照，对miR-219-5p与SOX5在个细胞系中的表达情况进行检测（RT-qPCR、Western blot）；并在SCC4及SCC9细胞株中验证miR-219-5p对SOX5表达的靶向调控作用。结果与正常人纤维细胞相比，miR-219-5p的表达水平显著降低，而SOX5的表达水平显著增加，差异有统计学意义（P < 0.05）；miR-219-5p与SOX5的表达水平具有负相关（P < 0.05）；在SCC4及SCC9细胞中，miR-219-5p通过结合3' 非编码区（3'-UTR）靶向调控SOX5的表达。结论 miR-219-5p可能通过对SOX5的靶向调控参与了口腔鳞状细胞癌的发生发展，为协助临床预后评估和/或筛选治疗的新候选药物提供了科学依据。
- 口腔癌细胞 /
- miR-219-5p /
- SOX5 /
- 靶向调控
Abstract: Objectives To explore whether the expressions of miR-219-5p and / or SRY-related high-mobility-group box 5 （SOX5） in oral squamous cell carcinoma cells are differ from normal cells. We also aim to confirm the transcription regulation in SOX5 by miR-219-5p. Methods Human oral squamous cell lines SCC4 and SCC9 were employed. The expression levels of miR-219-5p and SOX5 were detected in cancer cells （indicated by RT-qPCR and Western blot）. Genetic analysis and luciferase assay were employed to verify whether miR-219-5p targets SOX5 by binding to the specific region of 3'-UTR. Results Our data showed that high miR-219-5p and low SOX5 levels were detected in vitro SCC4 and SCC9 cells （P < 0.05）. miR-219-5p was negatively correlated with SOX5 expression. Genetic analysis and luciferase assay revealed that miR-291-5p regulated SOX5 expression by targeting the 3'untranslated region （3'-UTR） of the SOX5 mRNA. Conclusions This study demonstrates that miR-219-5p is involved in the development and prognosis of oral cancer by targeting the expression of SOX5, and can be a novel candidate for clinical prognosis and/or therapy.
- Oral cancer cells /
- miR-219-5p /
- SOX5 /
- Target regulation

HTML全文

图 1 miR-219-5p及SOX5在人口腔癌细胞株SCC4及SCC9中的表达检测

A：miR-219-5p在HF、SCC4及SCC9细胞株中的表达水平比较；B：SOX5 mRNA在HF、SCC4及SCC9细胞株中的相对表达水平；C：SOX5在HF、SCC4及SCC9各细胞株中的蛋白检测表达结果，左边为Western blot检测的印迹图，GAPDH为内参；右边为SOX5蛋白定量直方图。

Figure 1. The expressions of miR-219-5p and SOX5 in oral squamous cell lines SCC4 and SCC9

下载: 全尺寸图片幻灯片

图 2 miR-219-5p靶向3’-UTR特异性序列调控SOX5

A：miRMine预测人SOX5 mRNA序列中miR-219-5p可能的结合位点，红色标注的为miR-219-5p与SOX5 mRNA靶向结合的潜在位点；B：根据预测结果设计的miR-219-5p与SOX5 mRNA靶向结合潜在位点的突变序列（黄色标示）；3’UTR，3’非编码区；C：SCC4细胞的双荧光素酶报告实验结果；D：SCC9细胞的双荧光素酶报告实验结果。SOX5 3'UTR-wt，SOX5 3'UTR结合位点野生型组（未突变组）；SOX5 3'UTR-mut，SOX5 3'UTR结合位点突变组；miR-NC，miR-219-5p空白对照剂转染；miR-219-5p，miR-219-5p转染。

Figure 2. miR-219-5p regulates SOX5 by binding to specific sites in 3’-UTR

下载: 全尺寸图片幻灯片

图 3 在口腔癌细胞SCC4及SCC9中miR-219-5p靶向调控SOX5的表达

A：miR-219-5p在SCC4细胞中对SOX5 mRNA水平的表达调控； B：miR-219-5p在SCC9细胞中对SOX5 mRNA水平的表达调控；C：miR-219-5p在SCC4细胞中对SOX5蛋白水平的表达调控，上方为Western blot检测的蛋白印迹图，GAPDH为内对照，下方SOX5蛋白水平的定量统计直方图；D：miR-219-5p在SCC9细胞中对SOX5蛋白水平的表达调控，上方为Western blot检测的蛋白印迹图，GAPDH为内对照，下方SOX5蛋白水平的定量统计直方图。

Figure 3. miR-219-5p regulates the expression of SOX5 in SCC4 and SCC9

下载: 全尺寸图片幻灯片

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