华西口腔医学杂志 ›› 2021, Vol. 39 ›› Issue (1): 20-25.doi: 10.7518/hxkq.2021.01.003

• 基础研究 • 上一篇    下一篇

口腔白斑病癌变相关缺氧应答基因和微小RNA的芯片检测及表达验证

施琳俊1(), 杨溪2, 吴苏宁3, 刘伟2()   

  1. 1.上海交通大学医学院附属第九人民医院·口腔医学院口腔黏膜病科,国家口腔疾病 临床研究中心,上海市口腔医学重点实验室/上海市口腔医学研究所,上海 200011
    2.上海交通大学医学院附属第九人民医院·口腔医学院口腔颌面头颈肿瘤科,上海 200011
    3.江南大学附属医院(第三人民医院)口腔科,无锡 214000
  • 收稿日期:2020-03-20 修回日期:2020-11-21 出版日期:2021-02-01 发布日期:2021-03-02
  • 通讯作者: 刘伟 E-mail:shi-linjun@hotmail.com;liuweb@hotmail.com
  • 作者简介:施琳俊,副主任医师,博士,E-mail:shi-linjun@hotmail.com
  • 基金资助:
    国家自然科学基金(82074502);四川大学口腔疾病研究国家重点实验室开放课题(SKLOD2020OF08)

Transcriptome array screening and verification of oral leukoplakia carcinogenesis-related hypoxia-responsive gene and microRNA

Shi Linjun1(), Yang Xi2, Wu Suning3, Liu Wei2()   

  1. 1.Dept. of Oral Medicine, Shanghai Ninth People, s Hospital, Shanghai Jiao Tong University School of Medicine, National Clinical Research Center for Oral Diseases, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai 200011, China
    2.Dept. of Oral and Maxillofacial-Head and Neck Oncology, Shanghai Ninth People, s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
    3.Dept. of Stomatology, Affiliated Hospital of Jiangnan University/The Third People, s Hospital, Wuxi 214000, China
  • Received:2020-03-20 Revised:2020-11-21 Online:2021-02-01 Published:2021-03-02
  • Contact: Liu Wei E-mail:shi-linjun@hotmail.com;liuweb@hotmail.com
  • Supported by:
    The National Natural Science Foundation of China(82074502);Open Funding of the State Key Laboratory of Oral Diseases of Sichuan University(SKLOD2020OF08)

摘要: 目的

探讨口腔白斑病癌变进程中的缺氧应答基因及相关微小RNA (miRNA)的表达。

方法

利用Affymetrix GeneChip人转录组芯片(HTA)2.0对正常口腔黏膜、低危白斑、高危白斑、口腔早期鳞癌进行转录组学检测,基因本体论(GO)功能分析筛选出生物学作用为缺氧应答的基因和相关miRNA,并采用实时定量逆转录聚合酶链反应(qRT-PCR)检测验证白斑缺氧应答基因和miRNA的表达。

结果

正常黏膜与低危白斑间有7个缺氧应答差异基因,低危白斑与高危白斑间有10个缺氧应答差异基因,高危白斑与早期鳞癌间有21个缺氧应答差异基因。缺氧应答关键基因低氧诱导因子1α、趋化因子配体2、基质金属蛋白酶3 的mRNA和miR-21在正常黏膜、口腔白斑和早期鳞癌中的表达量均呈阶梯式升高,白斑与早期鳞癌之间的表达差异均具有统计学意义,这与转录组芯片结果基本一致。

结论

缺氧应答基因及相关miRNA参与白斑病癌变。

关键词: 口腔白斑病, 鳞状细胞癌, 缺氧应答基因, 微小RNA

Abstract: Objective

To study the hypoxia response gene and microRNA (miRNA) expression profiles in the pathogenesis and progression of oral leukoplakia (OLK).

Methods

Affymetrix GeneChip human transcriptome array 2.0 was used to detect the transcriptome of normal mucosa, low-risk OLK, high-risk OLK, and early squamous cell carcinoma (SCC). Gene ontology function analysis was used to screen genes and key miRNAs whose biological role is hypoxia response. Quantitative reverse transcription polymerase ch-ain reaction (qRT-PCR) was used to verify the expression of hypoxia response genes and miRNAs.

Results

A total of 7 different genes of hypoxia response between normal mucosa and low-risk OLK, 10 genes between low-risk and high-risk OLK, and 21 genes between high-risk OLK and SCC were identified. The results of qRT-PCR showed that the expression of hypoxia-inducible factor 1α, chemokine cc-motif ligand 2, and matrix metalloproteinase 3 mRNA and miR-21 in normal mucosa, OLK, and SCC increased in a stepwise manner. The expression difference between OLK and SCC was statistically significant and consistent with the results of transcriptome array.

Conclusion

The hypoxia response gene and related miRNA play roles in the development and progression of OLK.

Key words: oral leukoplakia, squamous cell carcinoma, hypoxia-responsive gene, microRNA

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