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Serbian Association for Cancer Research SDIRSACR

Autophagy 2023; 19: 1997-2014.
5. Patrick S, Lathoria K, Suri V, Sen E. Reduced YAP1 and FOLR1 in gliomas predict better response to chemotherapeutics.
Cell Signal 2023; 109: 110738.
6. Gowda P, Patrick S, Singh A, Sheikh T, Sen E. Mutant Isocitrate Dehydrogenase 1 Disrupts PKM2-beta-Catenin-BRG1
Transcriptional Network-Driven CD47 Expression. Mol Cell Biol 2018; 38.
7. Yang W, Soares J, Greninger P, Edelman EJ, Lightfoot H, Forbes S et al. Genomics of Drug Sensitivity in Cancer
(GDSC): a resource for therapeutic biomarker discovery in cancer cells. Nucleic acids research 2013; 41: D955-961.

L17

The (first) 20 years of BRCA testing in Croatia

Vesna Musani

Laboratory for hereditary cancer, Division of Molecular Medicine, Ruđer Bošković Institute, Zagreb, Croatia

Keywords: Breast neoplasms, Genetic Techniques, Genetic Testing, Hereditary Breast and Ovarian Cancer Syndrome,
Mutation, Ovarian neoplasms

Background: Hereditary Breast and Ovarian Cancer syndrome is primarily caused by inherited mutations in BRCA1 and
BRCA2 genes.
Patients and Methods: To establish a robust and quick detection method for BRCA1 and BRCA2 screening, healthy
women, over the age of 65, with no family history of cancer were tested for BRCA1 and BRCA2 variants to determine
the frequencies of harmless variants. 220 samples were screened for BRCA1 and 115 for BRCA2. Subsequently, 647
patients were tested for presence of deleterious mutations. Healthy controls and first 260 samples were analyzed by
HRM (high resolution melting) and Sanger sequencing for point mutations and QMPSF (quantitative multiplex PCR
of short fluorescent fragments) and LR-PCR (long-range PCR) for large rearrangements. The next 387 samples were
analyzed by NGS (next generation sequencing) and QMPSF, and results confirmed by Sanger sequencing and LR-PCR.
Results: Thirteen different BRCA1 mutations were found in 52 patients and fifteen different BRCA2 mutations were
found in 23 patients. Most common mutations were BRCA1 c.5266dupC (p.Gln1756fs) found in fifteen patients, BRCA1
c.181T>G (p.Cys61Gly) found in eleven patients, BRCA2 c.6641dupC (p.Tyr2215fs) found in eight patients and BRCA1
c.1252G>T (p.Glu418Ter) found in seven patients. In BRCA1 one large deletion of exons 4-6 was found in three non-
related patients, while in BRCA2 two large deletions were found in two patients, deletion of whole BRCA2 in one and
exons 1 and 2 in second.
Conclusions In Croatia: BRCA1 mutations are more frequent than BRCA2 mutations, with BRCA1 c.5266dupC being the
most common mutation. Large rearrangements are not common. Using HRM and QMPFS, or NGS and QMPFS to screen
for mutations and Sanger sequencing for confirmation are superior combination of methods for optimum results.

Acknowledgments and funding: 2005-2007 Technological project HITRA „Developing Method for Detection of Inherited
Predisposition to Breast Cancer in Croatia“, PI: Sonja Levanat, PhD
2009-2011 “Terry Fox Run” scientific project “Genetic Testing of Inherited Predisposition to Breast and Ovarian Cancer”,
PI: Sonja Levanat, PhD

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