Detection of genetic mutations in patients with breast cancer from Saudi Arabia using Ion AmpliSeq™ Cancer Hotspot Panel v.2.0

Abstract

Next-Generation Sequencing allows for quick and precise sequencing of multiple genes concurrently. Recently, this technology has been employed for the identification of novel gene mutations responsible for disease manifestation among breast cancer (BC) patients, the most common type of cancer amongst Arabian women, and the major cause of disease-associated death in women worldwide. Genomic DNA was extracted from the peripheral blood of 32 Saudi Arabian BC patients with histologically confirmed invasive BC stages I-III and IV, as well from 32 healthy Saudi Arabian women using a QIAamp ® DNA Mini Kit. The isolated DNA was quantified using a Qubit™ dsDNA BR Assay Kit with a Qubit 2.0 Fluorometer. Ion semiconductor sequencing technology with an Ion S5 System and AmpliSeq™ Cancer Hotspot Panel v2 were utilized to analyze ~2,800 mutations described in the Catalogue of Somatic Mutations in Cancer from 50 oncogenes and tumor suppressor genes. Ion Reporter Software v.5.6 was used to evaluate the genomic alterations in all the samples after alignment to the hg19 human reference genome. The results showed that out of the 50 genes, 26 mutations, including 17 (65%) missense point mutations (single nucleotide variants), and 9 (35%) frameshift (insertion/deletion) mutations, were identified in 11 genes across the cohort in 61 samples (95%). Mutations were predominantly focused on two genes, PIK3CA and TP53, in the BC genomes of the sample set. PIK3CA mutation, c.1173A>G located in exon 9, was identified in 15 patients (46.9%). The TP53 mutations detected were a missense mutation (c.215C>G) in 26 patients (86.70%) and 1 frameshift mutation (c.215_216insG) in 1 patient (3.33%), located within exon 3 and 5, respectively. This study revealed specific mutation profiles for every BC patient, Thus, the results showed that Ion Torrent DNA Sequencing technology may be a possible diagnostic and prognostic method for developing personalized therapy based on the patient's individual BC genome.

Utility of Circulating Cell-Free DNA in Assessing Microsatellite Instability and Loss of Heterozygosity in Breast Cancer Using Human Identification Approach

“…Breast cancer (BC) is the most common cancer in women, and is a leading cause of cancer-related deaths worldwide [1], including Saudi Arabia, with 18.7% of all cancer mortality in 2014, and 3629 new BC cases in 2018 reportedly affecting 14.8% of registered Saudi citizens [2]. BC is a complex, multifactorial disease, and is influenced by genetic and environmental factors including gender, age, hormones, obesity, BC family history, breastfeeding, and lifestyle [3,4]. As current BC screening focuses on detecting the associated genetic factors, including ATM (Ataxia Telangiectasia Mutated), CHEK2 (checkpoint kinase 2), Breast Cancer 1 (BRCA1), Breast Cancer 2 (BRCA2), and PALB2 (partner and localizer of BRCA2) [5], this necessitated the need for the discovery of BC biomarkers with sufficient diagnostic and prognostic sensitivity and specificity.…”

“…1 Student's t-test (2-tailed) for continuous variables, Pearson's χ 2 test for categorical variables 2. Mean ± SD 3. Number of subjects (percent total).…”

Tribulus terrestris Cytotoxicity against Breast Cancer MCF-7 and Lung Cancer A549 Cell Lines Is Mediated via Activation of Apoptosis, Caspase-3, DNA Degradation, and Suppressing Bcl-2 Activity

“…Multiple factors, such as hormonal, environmental, and hormonal factors, play a role in breast cancer development. The incidence of breast cancer is increasing in the Arab population, and a significant number of cases are diagnosed at the advanced stage of the disease, contributing to a higher mortality rate [5]. According to a Saudi Cancer Registry data on incidence from 2017, breast cancer was the most common cancer among women, and was responsible for 18.1% of all cancers.…”