LncRNA H19 Interaction with Mutant p53 Promotes Non-Small Cell Lung Cancer Progression

Abstract

Background: Long non-coding RNAs (lncRNA) regulate gene expression, while p53 is a tumor suppressor gene. Alterations in p53 expression could be important in lung carcinogenesis. This study explored the association and interaction of lncRNA and p53 in non-small cell lung cancer (NSCLC) development and progression. Methods: A eukaryotic expression vector carrying lncRNA H19 was constructed and transfected into NSCLC cells for the assessment of cell phenotype and gene expression through qRT-PCR, Western Blot, RNA-pull down, and immunoprecipitation-Western blot or qRT-PCR. Mutated p53 (p53mt) was assessed by using RT-PCR in H19-transfected tumor cells and 200 cases of NSCLC tissue samples for association with H19 expression. Results: p53 was over-expressed in NSCLC tissues, while H19 overexpression resulted in an increase in tumor cell growth and colony formation. H19 was able to promote p53-mediated transcriptional activity and H19 overexpression led to increased levels of p53 mRNA and protein. The RNA pull down-Western blot and immunoprecipitation-Western blot assays revealed the H19 was associated an increase in p53 expression. Ex vivo data showed that the p53 level was associated with advanced tumor-node-metastasis (TNM) stages and tobacco smoking or poor overall survival of patients. Conclusion: Both H19 and p53 are overexpressed in NSCLC tissues and their interaction promotes the p53 half-life and transcriptional activity, leading to NSCLC progression. Furthermore, high p53 and H19 expression is associated with poor overall survival of patients. Funding Statement: This study was supported in part by a grant from The Foundation of Medical Engineering Cross-work (#YG2016QN07). Declaration of Interests: The authors have no conflicts of interest to declare. Ethics Approval Statement: The study was approved by the Research Ethics Committee of Shanghai Jiaotong University (Shanghai, China) and written informed consent was obtained from all of the included patients.