Characterization of Alternatively Spliced Isoforms of MUC4 and ADAM12 Genes in a Metastatic Colorectal Cancer Cell Line Model

2021 ◽  
Author(s):  
Saleh Althenayyan ◽  
Mohammed H AlMuhanna ◽  
Abdulkareem Al Abdulrahman ◽  
Bandar Alghanem ◽  
Suliman A. Alsagaby ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Metastatic Colorectal Cancer ◽  
Cell Line ◽  
Differential Expression ◽  
Cell Lines ◽  
Cancer Cell ◽  
Colorectal Cancer Cell ◽  
Line Model ◽  
Cell Line Model ◽  
Alternatively Spliced

Colorectal cancer prognosis get worse with advancement of disease into metastatic stage. There is a pertinent need to develop prognostic biomarkers that can be used for personalized and precision medicine. Alternative splicing provides an insight into understanding of changes at isoform expression level which may not be evident at gene level. In this direction, we utilized our prior knowledge about significant alternatively spliced genes and chose ADAM12 and MUC4 for further characterization in a metastatic cell line model. These genes were found to be good prognostic indicators in The Cancer Genome Atlas database. We studied the gene organization and designed primers to specifically amplify a group of isoforms. Differential expression of these group of isoforms was observed in normal, primary and metastatic colorectal cancer cell lines. We further validated the results using sanger sequencing. Isoform expression was found to respond to the 5-fluorouracil treatment. RNAseq analysis of the cell lines further validated the differential expression of gene isoforms. Successful detection of ADAM12 and MUC4 in cell lysates varied according to the antibody used which may reflect differential expression of isoforms. This comprehensive study underscores the importance of studying alternatively spliced isoforms and their probable used as prognostic or predictive biomarkers.

scholarly journals Characterization of Alternatively Spliced Isoforms of MUC4 and ADAM12 Genes in a Metastatic Colorectal Cancer Cell Line Model

2021 ◽  
Author(s):  
Saleh Althenayyan ◽  
Mohammed H AlMuhanna ◽  
Abdulkareem Al Abdulrahman ◽  
Bandar Alghanem ◽  
Suliman A. Alsagaby ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Metastatic Colorectal Cancer ◽  
Cell Line ◽  
Differential Expression ◽  
Cell Lines ◽  
Cancer Cell ◽  
Colorectal Cancer Cell ◽  
Line Model ◽  
Cell Line Model ◽  
Alternatively Spliced

Colorectal cancer prognosis get worse with advancement of disease into metastatic stage. There is a pertinent need to develop prognostic biomarkers that can be used for personalized and precision medicine. Alternative splicing provides an insight into understanding of changes at isoform expression level which may not be evident at gene level. In this direction, we utilized our prior knowledge about significant alternatively spliced genes and chose ADAM12 and MUC4 for further characterization in a metastatic cell line model. These genes were found to be good prognostic indicators in The Cancer Genome Atlas database. We studied the gene organization and designed primers to specifically amplify a group of isoforms. Differential expression of these group of isoforms was observed in normal, primary and metastatic colorectal cancer cell lines. We further validated the results using sanger sequencing. Isoform expression was found to respond to the 5-fluorouracil treatment. RNAseq analysis of the cell lines further validated the differential expression of gene isoforms. Successful detection of ADAM12 and MUC4 in cell lysates varied according to the antibody used which may reflect differential expression of isoforms. This comprehensive study underscores the importance of studying alternatively spliced isoforms and their probable used as prognostic or predictive biomarkers.

scholarly journals Characterization of novel low passage primary and metastatic colorectal cancer cell lines

Oncotarget ◽  
2016 ◽  
Vol 7 (12) ◽  
pp. 14499-14509 ◽  
Author(s):  
Arnoud Boot ◽  
Jaap van Eendenburg ◽  
Stijn Crobach ◽  
Dina Ruano ◽  
Frank Speetjens ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Metastatic Colorectal Cancer ◽  
Cell Lines ◽  
Cancer Cell ◽  
Cancer Cell Lines ◽  
Colorectal Cancer Cell ◽  
Colorectal Cancer Cell Lines ◽  
Low Passage

Differential expression of small ubiquitin-like modifier family of proteins in patients with colorectal adenocarcinoma.

2011 ◽  
Vol 29 (4_suppl) ◽  
pp. 420-420
Author(s):  
R. Wiatrek ◽  
M. P. Duldulao ◽  
Z. Chen ◽  
W. Li ◽  
J. Kim ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Dna Repair ◽  
Differential Expression ◽  
Cell Lines ◽  
Cancer Cell ◽  
Expression Analysis ◽  
High Throughput ◽  
Colorectal Cancer Cell ◽  
Normal Tissues ◽  
Colorectal Cancer Cell Lines

420 Background: The cytotoxic effects of radiation and chemotherapy are mediated in part by DNA damage. The small ubiquitin-like modifier (SUMO) family of proteins regulates DNA repair pathways by activating factors involved in homologous recombination (HR) or non- homologous end joining (NHEJ). Our objective was to determine the potential role of SUMO proteins in patients with colorectal cancer. Methods: We first assessed two established colorectal cancer cell lines (HCT116 and HT29) and two normal colonic mucosa cell populations utilizing ultra high-throughput expression analysis (Solexa) to examine differential expression of genes involved in SUMOylation (SAE1, SAE2, PIAS-1, and DNAPKcs). To verify the clinical relevance of SUMOylation in colorectal cancer, we obtained archived specimens from patients with colorectal cancer (n=51) and examined the expression of these proteins in both benign and malignant tissue by immunohistochemistry (IHC). Results: Ultra high-throughput expression analysis of cancer and normal cells revealed a 10 to 15-fold upregulation of SAE2 (heterodimer of the E1 activation enzyme in SUMOylation), a 10 to 11-fold upregulation of DNAPKcs (catalytic subunit in NHEJ repair pathway), and a 6-fold upregulation of SAE1 in cancer cells. In contrast, PIAS-1 showed similar expression levels between cancer and normal colonic cells. By IHC, SAE1 and SAE2 were highly expressed in 65% and 53% of tumor specimens, respectively; but only in 7% and 0% of normal tissues, respectively. DNAPKcs was also highly expressed in tumor tissues (63%) with corresponding low expression (0%) in normal tissues. Corroborating the cell line results, PIAS-1 was infrequently expressed in both tumor (10%) and normal tissues (0%). Conclusions: We are the first to demonstrate the differential expression of SUMO proteins in colorectal cancer cell lines and in clinical specimens. SUMO proteins have a role in DNA repair and their expression in colorectal cancer may modify tumor response to chemoradiotherapy. No significant financial relationships to disclose.

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