Journal of Shanghai Jiao Tong University (Medical Science) >

2021 , Vol. 41 >Issue 3: 302 - 307

DOI: https://doi.org/10.3969/j.issn.1674-8115.2021.03.003

Basic research

Array detection and expression verification of DNA damage repair related genes in oral leukoplakia cancerization

  • Wei LIU ,
  • Min-wen ZHU ,
  • Lan WU
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  • 1.Department of Oral and Maxillofacial-Head and Neck Oncology, Shanghai Ninth People's Hospital, College of Stomatology, 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.Shanghai Xuhui District Dental Center, Shanghai 200032, China
    3.Department of Oral Mucosal Diseases, Shanghai Ninth People's Hospital, College of Stomatology, 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

Received date: 2020-03-30

  Online published: 2021-04-06

Supported by

National Nature Science Foundation of China(82074502);Innovative Research Team of High-Level Local Universities in Shanghai(SSMU-ZDCX20180901)

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Abstract

Objective

·To study the gene expression profile of DNA damage repair in the occurrence and development of oral leukoplakia (OL) and to verify the expression of mRNA and protein of key genes, ataxia-telangiectasia mutated (ATM) and histone variant H2A family member X (H2AX).

Methods

·Affymetrix HTA 2.0 array was used to detect the high-throughput transcriptome of normal mucosa (n=3), low-risk leukoplakia (n=4), high-risk leukoplakia (n=4) and early squamous cell carcinoma (n=6). Gene Ontology function analysis was used to screen out the genes related to DNA damage repair [|log2(chang fold, FC)|≥2.0 and P<0.05]. Real-time quantitative PCR (qPCR) and Western blotting were used to verify the mRNA and protein expression of key genes (ATM and H2AX) in human normal oral mucosal keratinocytes (HOK), OL (Leuk1) and squamous cell carcinoma cell lines (HN13), respectively.

Results

·DNA damage repair genes abnormally expressed in the occurrence and development of leukoplakia. There were 7 genes with |log2FC|≥2.0 from normal mucosa to low-risk leukoplakia, 7 genes with |log2FC|≥2.0 from low-risk leukoplakia to high-risk leukoplakia, and 52 genes with |log2FC|≥2.0 from high-risk leukoplakia to squamous cell carcinoma. qPCR revealed that the expression of ATM and H2AX in HOK cells, Leuk1 cells and HN13 cells increased stepwise (P<0.05). Western blotting revealed that the expression of γ-H2AX protein was also up-regulated in the HOK cells, Leuk1 cells and HN13 cells (P<0.05). ATM protein was up-regulated in Leuk1 cells compared with HOK cells, but down-regulated in HN13 cells (P<0.05).

Conclusion

·DNA damage repair genes, ATM and H2AX, are involved in the occurrence and development of OL.

Key words： oral leukoplakia (OL); squamous cell carcinoma; DNA damage repair; ataxia-telangiectasia mutated (ATM); histone variant H2A family member X (H2AX)

Cite this article

Wei LIU , Min-wen ZHU , Lan WU . Array detection and expression verification of DNA damage repair related genes in oral leukoplakia cancerization[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2021 , 41(3) : 302 -307 . DOI: 10.3969/j.issn.1674-8115.2021.03.003

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