Neoplasma. 2016;63(2):246-53. doi: 10.4149/210_150419N208.
Thevalueof SHOX2 methylation test in peripheralbloodsamplesusedforthedifferentialdiagnosisoflungcancerandotherlungdisorders.
Konecny M, Markus J, Waczulikova I, Dolesova L, Kozlova R, Repiska V, Novosadova H, Majer I.
Abstract
Methylationofthe cytosine residueswithintheCpGdinucleotidesplaysanimportant role in thefundamentalcellularprocesses, humandiseasesandevencancer. The DNA methylationrepresents averystable sign andthereforemaybeused as avaluablemarkerforcancerscreening. Epigeneticcancerbiomarkers are independent ofclassicalmorphologyandthus show extensivepotential to overcomethelimitationsof cytology. Severalepigeneticcancermarkershavebeenreported to bedetectable in body fluids such as bronchialaspirate, sputum, plasma andserum.Shortstaturehomeobox gene 2 (SHOX2) encodes ahomeo-domaintranscriptionfactor, which has beenidentified as aclosehomologueofthe SHOX gene andbothgenes are involved in skeletogenesisandheartdevelopment. Methylationof SHOX2 gene has beenshown to bepresentathigh prevalence in carcinomasoflung, howevermayalsobeused to identifyothertumourentities.Inthepresented study, wehavecomparedsuitabilityoftwotypesofmaterialassociatedwithlungcancerforthedetectionof SHOX2 methylation. Wehaveconfirmedthatmethylationof SHOX2 gene representsreliablemarkeroflungmalignancies. Theparalleltests in theblood plasma revealedthatitmayrepresent agoodalternativematerialfortestingofthe SHOX2 methylation, makingthe test available to patientswho are unable to undergobronchoscopy.
KEYWORDS:
DNA methylation test; SHOX2; blood plasma.; bronchiallavage; lungcancer
PMID:26774146
DOI:10.4149/210_150419N208
BreastCancer Res Treat. 2011 Feb;126(1):119-30. doi: 10.1007/s10549-010-1325-x. Epub 2011 Jan 4.
Comprehensivegeneticcharacterizationofhereditarybreast/ovariancancerfamiliesfromSlovakia.
Konecny M1, Milly M, Zavodna K, Weismanova E, Gregorova J, Mlkva I, Ilencikova D, Kausitz J, Bartosova Z.
Abstract
Germlinemutations in the BRCA1/2 genesaccountforthe majority ofhereditarybreastovariancancer (HBOC). Identificationofcausalmutationsmayhavesignificantimpact on clinical management of such families. Despitehighmutationdetectionrate, many HBOC casesremainwithoutidentified cause. These caseswarrant use ofseveralanalysismethods, such as thoseforlargegenomicrearrangementsand DNA copy numberchanges, oranalysisothergenes, shown to beassociatedwithincreased HBOC risk. Weassessed 585 Slovak HBOC forthe presence ofmutations in BRCA genes. Sequencingrevealedmutations in 100 families, representing 17.1% (88 and 12% ofmutationswerelocated in BRCA1 and BRCA2, respectively). Fourofthemutations, c.80+4del4, c.1938_1947del10 and c.1166delG in BRCA1 and c.6589delA in BRCA2 gene havebeendescribedonly in Slovakpopulation. Using MLPA analysis, wedetectedtwolargegenomicrearrangements in threefamilies, a deletionofexons 21 and 22, and a raredeletionof a whole BRCA1 gene. Twenty-sevendifferentvariantsofuncertainclinicaleffect (four novel) and 14 distinct SNP BRCA1 haplotypesweredetected. Theirpotentialeffectwasconsideredusingtheprediction software packagesAlign-GVGD, PmutandPolyphen. Weobservedthatthebestclinicalcriterionfortheinitiationof BRCA1 analysisisthe presence ofbreastcancerat 40 yearsofage in theassociationwiththe presence ofovariancancerdiagnosedaroundtheageof 50. Conversely, thebestclinicalcriterionforstartingwith BRCA2 analysisisthe presence ofbreastcancerdiagnosed in olderage (above 50), orthe presence ofbreastcancer in conjunctionwithcarcinomasatdifferentsitese.g., prostate, colorectum, ovary and uterus. Finallywehaveseenthattheanalysesofother HBOC risk gene TP53 andspecificmutation in CHEK2*c.1100delC in Slovak HBOC familieswere not efficientsince no mutationswerefound in these genes.
PMID:21203900
DOI:10.1007/s10549-010-1325-x
BreastCancer Res Treat. 2010 Jan;119(1):233-7. doi: 10.1007/s10549-008-0244-6. Epub 2008 Nov 15.
Identificationof a novel mutations BRCA1*c.80 + 3del4 and BRCA2*c.6589delA in Slovak HBOC families.
Konecny M1, Vizvaryova M, Zavodna K, Behulova R, GerykovaBujalkova M, Krivulcik T, Cisarik F, Kausitz J, Weismanova E.
Abstract
Mutations in the BRCA1 and BRCA2 genesaccountforthe majority ofhereditarybreastovariancancer (HBOC) cases. However, after BRCA1 and BRCA2 screeningstillthe most HBOC casesremain negative foranymutationalevent. Accordingly, in these casesraisesthe relevance to analyzetheunusual BRCA1/2 variantsofuncertainclinicalsignificance. Complex RNA/cDNAanalysismayconstitutethesolutionand help to interpret the HBOC syndrome in thefamily. In our study weanalyzedthe novel, to ourknowledge, not yetpublishedmutationsidentified in Slovak HBOC families, c.80 + 3del4 (IVS2 + 3delAGTC) in BRCA1 gene andmutation c.6589delA (6817delA) in BRCA2 gene. To determinetheeffectofthe BRCA1 mutation, weapplieddifferentapproaches: segregationanalysisofmutationwithdisease, presence in the set ofunaffectedcontrolsandfinally RNA/cDNA BRCA1 analysis. Novel BRCA2 mutationwasdeterminedperformingdirectsequencinganalysis. In conclusion, consideringtheresultsfromallusedtechniquesweapprovedthementionedmutations as seriouslypathogenicanddiseasecausingwithcleareffect on theonsetof HBOC syndrome.
PMID:19011960
DOI:10.1007/s10549-008-0244-6
BreastCancer Res Treat. 2008 Jun;109(3):581-3. Epub 2007 Jul 28.
Identificationofrarecomplete BRCA1 gene deletionusing a combinationof SNP haplotypeanalysis, MLPA andarray-CGH techniques.
Konecny M, Zavodna K, Vranova V, Vizvaryova M, Weismanova E, Mlkva I, Kuglik P, Kausitz J, Bartosova Z.
PMID:17661172
DOI:10.1007/s10549-007-9670-0
Neoplasma. 2007;54(2):137-42.
Thespectrumand incidence of BRCA1 pathogenicmutations in Slovakbreast/ovariancancerfamilies.
Konecny M1, Vizvaryova M, Weismanova E, Ilencikova D, Mlkva I, Weismann P, Machackova G, Kausitz J.
Abstract
Pathogenicgermlinemutations in BRCA1 and BRCA2 accountforthe majority ofhereditarybreast/ovariancancercases. Theanalysisof BRCA1 gene wascarriedout in 156 breast/ovariancancerfamilies: 82 familieswithstrongfamilyhistoryand 59 familieswith medium familyhistory. Generally, 31 familiesand 71 caseswith BRCA1 pathologicmutations (14 differenttypes) wereidentified in this study by combinationof SSCP anddirectsequencingtechniques. Usingapprovedsystematicnomenclaturenumbering, c.5266dupC (8 families, 21 cases), c.181T>G (5 families, 11 cases), c.68_69delAG (3 families, 5 samples) and c.843_846del4 (3 families, 4 samples) werethe most frequentlyfoundmutations in BRCA1 gene. Altogether these 4 mutationsaccountedfor 61.3% ofalldetectedpathogenicmutations in BRCA1. One novel mutation c.1166delG wasdetected in onefamily (4 cases). Frame-shiftmutationswerefound in 21 families (46 cases), nonsense mutations in 4 families (8 cases) andmissensemutations in 6 families (17 cases). Eventhoughthe 4 most frequentmutationsaccountfor 61.3% ofalldetectedpathogenicmutations, screeningofthewhole BRCA1 coding region isnecessary, due to thelargescaleoflowfrequencydiseasecausingmutations in breast/ovariancancerfamilies in Slovakia.
PMID:17319787