Next Article in Journal
The Effects of Astragalus membranaceus Active Extracts on Autophagy-Related Diseases
Previous Article in Journal
CAR-T with License to Kill Solid Tumors in Search of a Winning Strategy
Previous Article in Special Issue
Targeting of LRRC59 to the Endoplasmic Reticulum and the Inner Nuclear Membrane
Article Menu

Export Article

Open AccessArticle
Int. J. Mol. Sci. 2019, 20(8), 1902; https://doi.org/10.3390/ijms20081902

Quantification of the Lamin A/C Transcript Variants in Cancer Cell Lines by Targeted Absolute Quantitative Proteomics and Correlation with mRNA Expression

1
Department of Forensic Biology, Faculty of Forensic Sciences, Naif Arab University for Security Sciences, Riyadh 11452, Saudi Arabia
2
Department of Zoology, Faculty of Science, King Saud University, Riyadh 11362, Saudi Arabia
3
Newborn Screening & Biochemical Genetics lab, Department of Genetics, King Faisal Specialist Hospital and Research Center, Riyadh 11211, Saudi Arabia
4
Department of Biochemistry and Molecular Medicine, College of Medicine, Alfaisal University, Riyadh 11533, Saudi Arabia
5
Department of Chemistry, Memorial University of Newfoundland, St. John’s, NL A1B 3X7, Canada
*
Authors to whom correspondence should be addressed.
Received: 23 March 2019 / Revised: 11 April 2019 / Accepted: 15 April 2019 / Published: 17 April 2019
(This article belongs to the Special Issue Nuclear Envelope Proteins)
  |  
PDF [2927 KB, uploaded 17 April 2019]
  |  

Abstract

Lamin A/C proteins have key roles in nuclear structural integrity and chromosomal stability. Lamin A/C cumulative protein expression of all variants is reported by semi-quantitative Western blotting. To date, there have not been specific antibodies for the individual Lamin A/C transcript variants. We developed a mass spectrometric approach for the quantification of Lamin A/C transcript variants. A signature peptide for each specific splice variant of Lamin A/C was selected. A LC–MS/MS assay based on the selected signature peptides and their labeled internal standards was established to measure the expression of Lamin A/C transcript variant concentrations. The method validation was carried out according to Food and Drug Administration (FDA) guidelines. The expression levels of the Lamin A/C transcript variants were measured in samples derived from MCF7 and U937 cell lines. RT-qPCR assay was also used to quantitate and compare the mRNA expression of splice variants of Lamin A/C. The established and validated method showed a great linearity, sensitivity, and precision. The different expressed Lamin A/C variants in different cell lines were measured and their levels were in concordance with qRT-PCR results. The developed method is reproducible, reliable, and sensitive for measuring different Lamin A/C transcript variants in different cell lines. View Full-Text
Keywords: lamin A; lamin C; lamin AΔ10; lamin AΔ50 (Progerin); LC–MS/MS; MCF7; U937 lamin A; lamin C; lamin AΔ10; lamin AΔ50 (Progerin); LC–MS/MS; MCF7; U937
Figures

Graphical abstract

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
SciFeed

Share & Cite This Article

MDPI and ACS Style

Al-Qahtani, W.S.; Abduljabbar, M.; AlSuhaibani, E.S.; Abel Rahman, A.; Aljada, A. Quantification of the Lamin A/C Transcript Variants in Cancer Cell Lines by Targeted Absolute Quantitative Proteomics and Correlation with mRNA Expression. Int. J. Mol. Sci. 2019, 20, 1902.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top