Silencing NID2 by DNA Hypermethylation Promotes Lung Cancer
To characterize the DNA methylation as well as exploring the relationship between NID2 methylation and the lung cancer development. Collecting chip data of 9 lung cancer samples and 11 adjacent normal samples from the Gene Expression Omnibus database. Tissues and cells NID2 gene methylation level was measured by methylation-specific PCR. NID2 mRNA level and protein level were validated by Real-Time PCR and Western blot separately. Functional study of lung cancer cells was performed with Cell Counting Kit-8 assay. Colony formation assay, transwell assay, wound healing assay and low cytometry were performed. Finally, NID2 tumorigenesis in vivo was tested in nude mice xenograft models. Microarray analysis outcome present NID2 hypermethylation status in lung cancer tissues. High methylation and low mRNA expression levels of NID2 were detected. After NID2 demethylation or overexpression in cancer cells, cell viability, proliferation, migration as well as invasion ability decreased. Nevertheless, a significant enhancement in apoptosis rate were observed. Overexpressing NID2 or demethylation in lung cancer cells inhibited the tumorigenesis of lung cancer in nude mice. The mRNA and protein level of NID2 in tumors obtained from nude mice xenograft were unanimous with the in vitro assays’ outcome, which significantly decreased after overexpressing NID2 or demethylation. NID2 methylation reduces its expression level and promotes the development of lung cancer.
KeywordsLung cancer DNA methylation NID2 Nude mice xenograft
Substantial contribution to the conception and design of the work: Jianfeng Wang, Yan Zhao and Hongyan Xu; Acquisition, analysis, and interpretation of the data: Jianfeng Wang, Jun Ma, Feihai Liang and Qingxu Zou; Drafting the manuscript: Jianfeng Wang; Revising the work critically: Fengwu Lin; Final approval of the the version to be published: All authors.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
All procedures performed in studies involving human participants were in accordance with the ethical standards of the China-Japan Union Hospital of Jilin University committee. Informed consent was obtained from all individual participants included in the study.
All procedures involving animals were performed in compliance with guidelines of Q China-Japan Union Hospital of Jilin University.
Informed consent was obtained from all individual participants included in the study.
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