Validation of the methylation level of two long noncoding RNAs in non-small cell lung and ovarian cancer patients

Urraca Arias, Ruth Itziar (2019). Validation of the methylation level of two long noncoding RNAs in non-small cell lung and ovarian cancer patients. Proyecto Fin de Carrera / Trabajo Fin de Grado, E.T.S. de Ingeniería Agronómica, Alimentaria y de Biosistemas (UPM), Madrid.

Description

Title: Validation of the methylation level of two long noncoding RNAs in non-small cell lung and ovarian cancer patients
Author/s:
  • Urraca Arias, Ruth Itziar
Contributor/s:
  • Orellana Saavedra, Juan
  • Ibáñez de Cáceres, Inmaculada
Item Type: Final Project
Degree: Grado en Biotecnología
Date: June 2019
Subjects:
Faculty: E.T.S. de Ingeniería Agronómica, Alimentaria y de Biosistemas (UPM)
Department: Biotecnología - Biología Vegetal
Creative Commons Licenses: Recognition - No derivative works - Non commercial

Full text

[img] PDF - Users in campus UPM only - Requires a PDF viewer, such as GSview, Xpdf or Adobe Acrobat Reader
Download (3MB)

Abstract

Epigenetics has emerged as an important research area in many diseases, including cancer. Specifically, DNA methylation is the best-known epigenetic footprint that regulates gene expression. Its implication in cancer has been widely described, particularly as a general hypomethylation and a specific hypermethylation in tumor suppressor genes. Noncoding RNAs (ncRNAs) profiles are other epigenetic elements involved in gene expression as well. Cancer research focus on these ncRNAs has increased in the past years as well as its relation with chemotherapy resistance, reporting several advances. Among all types of ncRNAs, this study focused on long noncoding RNAs (lncRNAs), combined with the analysis of DNA methylation in the promoter regions. Based on previous findings from an epigenomic study carried out at our laboratory, two lncRNAs were selected for the present work: AC141928.1 and AC091814, which were identified as differentially methylated in resistant phenotypes from eight paired ovarian cancer and NSCLC cell lines, respectively. The quantitative validation of the methylation levels of both lncRNAs in the ovarian cancer cell lines OVCAR3 and A2780 for AC141928.1 and lung cancer cell lines H23 and H460 for AC091814, was carried out through quantitative Methylation Specific PCR (qMSP), confirming previous results obtained by using a semiquantitative technique (Bisulfite Sequencing, BS) from the mentioned epigenomic study: OVCAR3 and H23 presented differential methylation when comparing the cisplatin-sensitive and resistant variants. Afterwards, a translational approach was conducted to validate results from cellular models in cancer patients and normal donors. On one hand, 10 control samples from healthy tissue obtained from tubal ligations presented unmethylated CpG positions (BS) for AC141928.1 with a low quantitative average methylation level of 10.29%, while 48 tissue samples from ovarian cancer samples showed diverse methylation levels. In this way, a tendency was found towards patients with lower methylation levels and higher survival. On the other hand, two non-tumor control samples for AC091814 exhibited mainly hemimethylated CpG positions (BS) with a high average methylation level of 90.57%, while two additional groups of control saliva samples from 9 high-risk smokers (HRS) and 6 Chronic Obstructive Pulmonary Disease (COPD) patients presented lower methylation levels of 70.67% and 63.04%, respectively. 21 NSCLC tissue samples did not show significant differences in methylation compared to HRS and COPD patients (72.23%). In conclusion, the hypermethylation of the lncRNA AC141928.1 may be a potential prognostic biomarker for high-risk ovarian cancer patients in response to cisplatin-based chemotherapy, which needs to be confirmed by an extended cohort of ovarian cancer patients, while AC091814 promoter demethylation could be implicated in the early lung tumor establishment. This study has also confirmed the importance of validating the results obtained from cell lines, in control and pathogenic samples from patients, before the development of any confident translational approach.

More information

Item ID: 57586
DC Identifier: http://oa.upm.es/57586/
OAI Identifier: oai:oa.upm.es:57586
Deposited by: Biblioteca ETSI Agrónomos
Deposited on: 24 Dec 2019 07:44
Last Modified: 24 Dec 2019 07:44
  • Logo InvestigaM (UPM)
  • Logo GEOUP4
  • Logo Open Access
  • Open Access
  • Logo Sherpa/Romeo
    Check whether the anglo-saxon journal in which you have published an article allows you to also publish it under open access.
  • Logo Dulcinea
    Check whether the spanish journal in which you have published an article allows you to also publish it under open access.
  • Logo de Recolecta
  • Logo del Observatorio I+D+i UPM
  • Logo de OpenCourseWare UPM