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作者 Stella S, Vitale SR, Martorana F, Massimino M, Pavone G, Lanzafame K, Bianca S, Barone C, Gorgone C, Fichera M, Manzella L
Stefania Stella, 1,2 Silvia Rita Vitale, 1,2 Federica Martorana, 1,2 Michele Massimino, 1,2 Giuliana Pavone, 3 Katia Lanzafame, 3 Sebastiano Bianca, 4 Chiara Barone, 5 Cristina Gorgone, 6 Marco Fichera 2 Explorer University, 16 Umtholampilo wase-Capital Lichera1, I-Maperitanial University, 16 Umtholampilo we-Caperitanial1 , Catania, 95123, Italy;2 Centre for Experimental Oncology and Hematology, AOU Policlinico “G.Rodolico – San Marco”, Catania , 95123, Italy;3 I-Medical Oncology, i-AOU Policlinico “G.Rodolico – San Marco”, Catania, 95123, Italy;4 Medical Genetics, ARNAS Garibaldi, Catania, 95123, Italy;5 Medicine Genetics, ASP, Syracuse, 96100, Italy;6 UMnyango Wezesayensi Yezinto Eziphilayo kanye Nezobuchwepheshe, eNyuvesi yaseCatania, i-Medical Genetics, Catania, Italy, 95123;I-7Oasi Research Institute-IRCS, Troina, 94018, Italy Ezokuxhumana: Stefania Stella, ucingo +39 095 378 1946, i-imeyili [i-imeyili ivikelwe];[i-imeyili ivikelwe] Injongo: Ukuguqulwa kwe-germline ku-BRCA1 ne-BRCA2 kanye nokusungula umdlavuza webele (BC), i-ovary (OC) nokunye okuhlotshaniswa nobungozi bokuphila impilo yonke yomdlavuza.Ukuhlola isakhi sofuzo se-BRCA kuyisihluthulelo sokuhlola ubungozi bomuntu ngamunye, kanye nokuthola izindlela zokuvimbela kubathwali abanempilo kanye nokuhlanganisa ukwelashwa ezigulini ezinomdlavuza. Izinhlobonhlobo ze-BRCA ze-pathogenic emindenini yaseSicily, izifundo eziqondiswe ngokuqondile kubantu empumalanga yeSicily ziyashoda.Inhloso yocwaningo lwethu kwakuwukuphenya izehlakalo nokusatshalaliswa kwe-BRCA pathogenic germline alterations eqenjini leziguli ze-BC ezivela empumalanga yeSicily kanye nokuhlola ukuhlotshaniswa kwazo nezici ezithile ze-BC kusetshenziswa ukushintshwa kwesizukulwane esilandelayo sokulandelana kwe-tumor kanye ne-3. Iziguli ezingu-5 (9%) zibe nokuhluka kwe-BRCA pathogenic, 17 (49%) ku-BRCA1 kanye ne-18 (51%) ku-BRCA2.BRCA1 izinguquko zivame kakhulu ezigulini ze-BC ezintathu ezimbi, kuyilapho ukuguqulwa kwe-BRCA2 kuvame kakhulu ezigulini ze-luminal BC.Uma kuqhathaniswa nabangewona abathwali, izifundo ezitholakalayo ezinenkomba ephezulu ye-BRCA1 zinikeza ngokuphawulekayo i-tumor1. Isimo sokuguqulwa kwe-BRCA ezigulini ze-BC ezivela empumalanga ye-Sicily futhi siqinisekisa indima yokuhlaziywa kwe-NGS ekuhlonzeni iziguli ezine-BC. Sekukonke, le datha ihambisana nobufakazi bangaphambilini obusekela ukuhlolwa kwe-BRCA ukuze kuvinjwe indlela efanele kanye nokwelashwa komdlavuza kubathwali abaguqukayo.
Umdlavuza webele (BC) uyisifo esibulalayo esivame kakhulu emhlabeni wonke kanye nomdlavuza obulala kakhulu kwabesifazane.1 Izici zezinto eziphilayo ezinquma ukubikezelwa kwe-BC kanye nokuziphatha komtholampilo kuye kwacwaningwa kabanzi futhi kwacaciswa kancane ngokuhamba kwesikhathi.Eqinisweni, omaka abambalwa be-surrogate okwamanje basetshenziselwa ukuhlukanisa i-BC ibe yizinhlobo ezahlukene zama-molecular subtypes.Ziyi-estrogen (ER) kanye/noma i-progesterone yokukhula komuntu, i-progesterogen yomuntu (i-progesterogen2), i-progesterone yomuntu inkomba yesilinganiso se-Ki-67 kanye nebanga lesimila (G) .2 Ukuhlanganiswa kwalezi ziguquguqukayo kuhlonze izigaba ze-BC ezilandelayo: 1) Izimila ze-Luminal, ezibonisa i-ER kanye/noma i-PgR expression, zibalelwa ku-75% we-BCs. Lezi zicubu zaphinde zahlukaniswa zaba i-Luminal A, lapho i-Ki-67 ingaphansi kuka-20% kanye ne-HER2 negative, kanti i-Luminal B, ilingana ne-0% ye-Ki-6 noma i-2plification ye-prolife2 ku-HER2% noma i-prolife2. inkomba;2) amathumba e-HER2+ ane-ER ne-PgR angenayo kodwa abonisa ukukhuliswa kwe-HER2.Leli qembu lihlanganisa u-10% wazo zonke izimila zamabele;I-3) Umdlavuza webele we-Triple-negative (TNBC), ongabonisi inkulumo ye-ER ne-PgR kanye nokukhulisa i-HER2, i-akhawunti cishe ye-15% yomdlavuza webele.2-4
Phakathi kwalezi zinhlobo ezincane ze-BC, ibanga le-tumor kanye ne-proliferation index limelela ama-biomarker e-cross-sectional ahlotshaniswa ngokuqondile nokuzimela nokuhlukunyezwa kwe-tumor kanye ne-prognosis.5,6
Ngaphandle kwezici zezinto eziphilayo ezishiwo ngenhla, indima yokuguqulwa kwezakhi zofuzo eziholela ekuthuthukisweni kwe-BC ibaluleke kakhulu eminyakeni embalwa edlule.7 Cishe i-1 kwezingu-10 izimila zesifuba zitholwa njengefa ngenxa yokuguqulwa kwamagciwane ezakhini ezithile zofuzo.8 Izifundo ezimbili ezinkulu ze-epidemiological ezihilela ngaphezu kwe-180,000 abesifazane abasanda kuhlonza i-genes, i-18 TM, i-1BR, i-BCA, i-10, i-8, i-B. I-K2, i-PALB2, i-RAD51C, ne-RAD51D) ngokuyinhloko inesibopho sofuzo lwe-BC.Phakathi kwalezi zakhi zofuzo, i-BRCA1 ne-BRCA2 (ngemuva kwalokho ebizwa ngokuthi i-BRCA1 / 2) ibonise ukuhlobana okunamandla kakhulu nokuthuthukiswa kwezicubu zesifuba. I-ectal, ne-melanoma.Kusukela eminyakeni eyi-13 kuye kwengama-80, izehlakalo ezikhulayo ze-BC zingama-72% kwabesifazane abane-BRCA1 pathogenic variant (PV) kanye nama-69% kwabesifazane abane-BRCA2 PV.14
Ngokuphawulekayo, ukushicilelwa kwakamuva kusikisela ukuthi ingozi ye-BC incike ohlotsheni lwe-PV.Eqinisweni, uma kuqhathaniswa nezinhlobonhlobo ze-pathogenic truncating, ukuhlukahluka kwe-glaring missense, ikakhulukazi esakhiweni se-BRCA1, kuhlotshaniswa nengozi encishisiwe ye-BC, ikakhulukazi kwabesifazane asebekhulile.15
Ukuba khona kwe-BRCA1 noma i-BRCA2 PV kwakuhlotshaniswa nezici ezihlukahlukene zebhayoloji nezokwelapha. izicubu ze-se zivame kakhulu ku-lumen B futhi ngokuvamile zenzeka kubantu abadala asebekhulile.16-18 Ngokuphawulekayo, ukuguqulwa kwezinguquko ku-BRCA1 ne-BRCA2 kwandisa ukuzwela ekwelapheni okuthile, kuhlanganise nosawoti we-platinum nezidakamizwa ezihlosiwe ezifana ne-poly(ADP-ribose) polymerase inhibitors (PARPi) .19,20
Eminyakeni embalwa edlule, ukuqaliswa kokulandelana kwesizukulwane esilandelayo (NGS) ekusebenzeni komtholampilo kuye kwanika amandla inani elandayo leziguli ze-BC ukuthi zihlolwe amangqamuzana ezimo ezithinta umdlavuza, okuhlanganisa i-BRCA1/2.21 Ngasikhathi sinye, izincazelo ezisekelwe ezimisweni eziqondile eziphathelene nomlando womndeni , izibalo zabantu, kanye nezici ze-clinicopathological ukuze kuhlonzwe kangcono abantu ngabanye23c 23 ukuhlolwa komtholampilo. Ukuhlolwa kwe-BRCA1/2 kubantu abathile, okugqamisa umehluko ezifundeni zezwe.24–27 Nakuba kukhona imibiko ngeqoqo le-BC entshonalanga yeSicily, idatha embalwa iyatholakala ekuhlolweni kwe-BRCA1/2 kubantu baseSicily empumalanga.28,29
Sichaza lapha imiphumela yokuhlolwa kwe-germline BRCA1/2 ezigulini ze-BC ezivela empumalanga yeSicily, siphinde sihlobanise ubukhona bezinguquko ze-BRCA1 noma ze-BRCA2 nezici eziyinhloko ze-clinicopathological zalezi zimila.
Ucwaningo lwe-retrospective lwenziwa "Esikhungweni Se-Experimental Oncology and Hematology" esibhedlela sase-Policlinico.Rodolico - San Marco eCatania.Kusukela ngoJanuwari 2017 kuya ku-March 2021, ingqikithi yeziguli ezingu-455 ezinomdlavuza webele kanye ne-ovarian, i-melanoma, i-pancreatic noma i-prostate zadluliselwa ocwaningweni lwethu lwe-molecular diagnostic / BRCAratory ukuhlolwa kofuzo. lsinki, futhi bonke ababambiqhaza banikeze imvume ebhaliwe enolwazi ngaphambi kokuhlaziywa kwamangqamuzana.
Izici ze-Histological and Biological (ER, PgR, HER2 status, Ki-67, and grade) of BC ziye zahlolwa kumasampuli ayinhloko e-biopsy noma okuhlinzwa, kucatshangelwa izingxenye zesimila ezinolaka kuphela.Ngokusekelwe kulezi zici, ama-BC ahlukaniswe ngale ndlela elandelayo: i-luminal A (ER+ kanye/noma i-PGR+, HER2-, (ER20), i-HER2-, i-HER20, i-HER20, i-luminal, i-HER2, i-HER2, i-HER20, i-6, i-7 -67≥20%), i-luminal B-HER2+ (ER kanye/noma i-PgR+, HER2+), HER2+ (ER ne-PgR-, HER2+) noma i-negetive kathathu (ER ne-PgR-, HER2-).
Ngaphambi kokuhlola isimo sokuguqulwa kwe-BRCA1 kanye ne-BRCA2, ithimba lezinhlaka eziningi okuhlanganisa i-oncologist, isazi sofuzo, kanye nodokotela wezengqondo benze ukubonisana kofuzo lwesimila esigulini ngasinye ukuze kutholwe ubukhona be-BRCA1 kanye/noma i-BRCA1.noma abantu abasengozini enkulu ye-PV kufuzo lwe-BRCA2.Ukukhethwa kwesiguli kwenziwa ngokuvumelana neziqondiso ze-Italian Society of Medical Oncology (AIOM) kanye nezincomo zendawo zase-Sicilian.30,31 Lezi zindlela zifaka: (i) umlando womndeni wezinhlobonhlobo ezaziwayo ze-pathogenic ezakhini zofuzo ezithintekayo (isb, BRCA1, BRCA2, TP53, PTEN);(ii) abesilisa abanoBC;(iii) labo abano-BC no-OC;(iv) abesifazane abane-BC weminyaka engama-50 kanye nomlando womndeni ka-BC, OC, noma umdlavuza we-pancreatic wezihlobo eziyizihlobo zokuqala komunye nomunye (kuhlanganise nezihlobo ayizihlobo ze-first degree);(vii) Umlando womuntu siqu ka-OC kanye okungenani nesihlobo esisodwa sezinga lokuqala: (a) BC Kutholwe isampula yegazi elingu-20 mL esigulini ngasinye futhi laqoqwa kumashubhu e-EDTA (BD Biosciences).I-Genomic DNA yahlukaniswa kusukela kumasampula egazi aphelele angu-0.7 mL kusetshenziswa i-QIAsymphony DSP DNA Midi kit Isolation Kit (QIAGEN, Hilden, Italy) ngokuvumelana neziqondiso zomkhiqizi futhi yadlula ku-3. , MA, USA) Yenza ukulinganisa.Ukulungiswa okuhlosiwe kanye nokulungiswa komtapo wezincwadi kwenziwa umpheki we-Oncomine™ BRCA Research Assay Chef, olungele ukulayishwa ku-Ion AmpliSeq™ Chef Reagents DL8 Kit ukuze alungiselele umtapo wezincwadi ngokuzenzakalelayo ngokuya ngemiyalelo yomkhiqizi. Ikhithi iqukethe amachibi amabili e-PCR e-multiplex angasetshenziswa ukutadisha yonke i-BRCA7 (N500030) (N5M0030). ) izakhi zofuzo. Kafushane, u-15 µL wesampula ngayinye ehlanjululiwe ye-DNA (10 ng) yengezwe ezingcwecweni ezinebhakhodi ukuze kulungiselelwe umtapo wolwazi futhi zonke izinto ezisebenzayo nezinto ezisetshenziswayo zalayishwa ethuluzini le-Ion Chef™. Ukulungiswa komtapo wolwazi okuzenzakalelayo kanye nokuhlanganisa amasampula omtapo wezincwadi anebhakhodi kwase kwenziwa ethuluzini le-Ion Chef™. cientific, Waltham, MA, USA) ngokwemiyalelo yomkhiqizi. Ekugcineni, imitapo yolwazi ihlanganiswa ngezilinganiso ezilinganayo kumashubhu esampula omtapo wezincwadi we-Ion Chef™ (amashubhu anebhakhodi) futhi alayishwa kuthuluzi le-Ion Chef™. Ukulandelanisa kwenziwa kusetshenziswa i-Ion Torrent S5 (Thermo Fisher Scientific) kusetshenziswa ithuluzi le-Ion Fisher Scientific (I-Fisher) Isayensi).Ukuhlaziywa kwedatha kwenziwa yi-Amplicon Suite (SmartSeq srl) kanye ne-Ion Reporter Software.
Zonke izinhlobo zegama ezihlukile zilandela imihlahlandlela yamanje ye-Human Genome Variation Consortium, etholakala ku-inthanethi (HGVS, http://www.hgvs.org/mutnomen).Ukubaluleka komtholampilo kokuhlukahluka kwe-BRCA1/2 kwachazwa kusetshenziswa ukuhlukaniswa kwe-International Consortium ENIGMA (Inethiwekhi Esekelwe Ebufakazini Ye-Mutanteniing Alemconsor/Isizindalwazi se-Mutantening/org) njenge-ARUP, BRCAEXCHANGE , ClinVar, IARC_LOVD, kanye ne-UMD. Ukuhlukaniswa kuhlanganisa izigaba ezinhlanu ezihlukene zengcuphe: i-benign (isigaba I), okungenzeka ibenign (isigaba II), okuhlukile kokubaluleka okungaqinisekile (VUS, isigaba III), okungenzeka kube yi-pathogenic (isigaba IV), kanye ne-funcogenicity (isigaba se-protein esishintshashintshayo kanye nomphumela we-analogue of V). ukufinyelela kuma-database angama-30.32
Ukunikeza ukubaluleka komtholampilo okungaba khona ku-VUS ngayinye, kusetshenziswe ama-algorithms wokubikezela amaprotheni ekhompyutha alandelayo: UKUGUQUKA TASTER, 33 PROVEAN-SIFT (http://provean.jcvi.org/index.php), POLYPHEN-2 (http:// /genetics.bwh.harvard.edu/pph2/G) kanye ne-Align. d_input.php).Okuhlukile okufakwe esigabeni 1 no-2 kwakubhekwa njengohlobo lwasendle.
Ukulandelana kwe-Sanger kuqinisekise ubukhona bokwehluka ngakunye kwe-pathogenic. Kafushane, ipheya yeziqalo ezithile zadizayinelwa ukwahluka ngakunye okutholiwe ngokusebenzisa i-BRCA1 kanye ne-BRCA2 yokulandelanisa kofuzo kwereferensi (NG_005905.2, NM_007294.3 kanye ne-NG_012772.3, NM5772.3, NM9. nciza.
Iziguli ezitholwe zingenayo i-BRCA1/2 gene zihlolwe i-multiplex ligation-dependent probe amplification (MLPA) ngokuya ngemiyalo yomkhiqizi yokuhlola ukuba khona kokuhlelwa kabusha kwe-genomic (LGR). ama-nucleotide ngobude.Imikhiqizo yokukhulisa i-Probe, ehlanganisa isethi eyingqayizivele yamaamplikhoni e-PCR, yabe isihlaziywa nge-capillary electrophoresis kanye nesofthiwe ye-Cofalyser.Net ngokuhlanganyela namatafula afanele e-batch e-Cofalyser (www.mrcholland.com).
Okuguquguqukayo okukhethiwe kwe-clinicopathological (ibanga le-histological kanye ne-Ki-67% proliferation index) kuhlotshaniswa nokuba khona kwe-BRCA1/2 PV, kubalwa kusetshenziswa isofthiwe ye-Prism v. 8.4 kusetshenziswa ukuhlolwa okuyingqophamlando kukaFisher kucatshangwa ukuthi inani le-p <0.05 libalulekile.
Phakathi kukaJanuwari 2017 kanye noMashi 2021, iziguli ezingu-455 zahlolelwa ukuguqulwa kwegciwane le-BRCA1/2. Ukuhlolwa kokuguqulwa kwe-germline kwenziwa Esikhungweni Sesibhedlela Se-Policlinico Se-Experimental Oncology and Hematology. Ngokuvumelana nesiqondiso sase-Sicilian (http://www.gurs.regione.sicilia.sicilia.it/Indicep. 2020), iRodolico yaseCatania - San Marco” isiyonke, iziguli ezingama-389 Kube nomdlavuza webele, umdlavuza wamabele angama-37, umdlavuza wamapancreas awu-16, umdlavuza we-prostate oyisi-8 kanye ne-melanoma emi-5.Ukusatshalaliswa kweziguli ngokohlobo lomdlavuza kanye nemiphumela yokuhlaziya kuboniswa kuMfanekiso 1.
Umfanekiso 1 ubonisa ishadi eligelezayo elibonisa ukubuka konke kocwaningo.Iziguli ezinebele, i-melanoma, i-pancreatic, i-prostate, noma izimila ze-ovarian zahlolelwa ukuguqulwa kwezakhi zofuzo ze-BRCA1 ne-BRCA2.
Izifinyezo: ama-PV, ukuhluka kwe-pathogenic;I-VUS, okuhlukile kokubaluleka okungaqinisekile;I-WT, ukulandelana kohlobo lwasendle lwe-BRCA1/2.
Sigxile kakhulu ezifundweni zethu kumaqoqo omdlavuza webele.Iziguli zazineminyaka yobudala ephakathi kweminyaka engu-49 (ububanzi obungu-23-89) futhi kwakungabesifazane kakhulu (n=376, noma 97%).
Kulezi zihloko, ama-64 (17%) abe nokuguqulwa kwe-BRCA1/2 futhi bonke bekungabantu besifazane.Amashumi amathathu nanhlanu (9%) ane-PV futhi angu-29 (7.5%) ane-VUS.Ishumi nesikhombisa (48.6%) lokuhlukahluka okungu-35 kwe-pathogenic kwenzeke ku-BRCA1 no-18 (51.4%) ku-BRCA2, kuyilapho u-51.4%) ku-. (Amanani 1 kanye no-2).I-LGR yayingekho ekuhlaziyeni kwe-MLPA.
Umfanekiso 2. Ukuhlaziywa kwezinguquko ze-BRCA1 kanye ne-BRCA2 ezigulini zomdlavuza webele we-389. (A) Ukusatshalaliswa kwezinhlobonhlobo ze-pathogenic (PV) (obomvu), ukuhlukahluka kokubaluleka okungaqinisekile (VUS) (iwolintshi), kanye ne-WT (eluhlaza okwesibhakabhaka) ezigulini zomdlavuza webele we-389;(B) Iziguli ze-389 zomdlavuza webele Amashumi amathathu nanhlanu (9%) ayene-BRCA1 / 2 variants pathogenic (PVs) .Phakathi kwabo, i-17 (48.6%) yayiyi-BRCA1 PV carriers (ibomvu emnyama) futhi i-18 (51.4%) yayingabathwali be-BRCA2 (obomvu okhanyayo);(C) Ama-29 (7.5%) wezifundo ezingama-389 aphathe i-VUS, 5 (17.2%) i-BRCA1 yofuzo (iwolintshi elimnyama) kanye nama-24 (82.8%) wofuzo we-BRCA2 (owolintshi okhanyayo).
Izifinyezo: ama-PV, ukuhluka kwe-pathogenic;I-VUS, okuhlukile kokubaluleka okungaqinisekile;I-WT, ukulandelana kohlobo lwasendle lwe-BRCA1/2.
Ngokulandelayo siphenye ukusabalala kwe-BC molecular subtypes ezigulini ezine-BRCA1/2 PV. Ukusatshalaliswa kufaka phakathi i-luminal A engu-2 (5.7%), 15 (42.9%) luminal B, 3 (8.6%) luminal B-HER2+, 2 (5.7%) HER2+ kanye no-13 (37.1%) phakathi kweziguli ezine-luminal TN B. , 2 (11.8%) babenesifo se-HER2+, futhi abangu-10 (58.8%) babene-TNBC.Amathumba angenawo ukuguqulwa kwe-BRCA1 kwakungase kube yi-luminal A noma i-luminal B-HER2+ (Figure 3).Eqenjini elingaphansi le-BRCA2-positive, 10 (55.6%) izimila zaziyi-luminal B, 3 (16) , 16 BBC (16%) kanye ne-BTN 16%. .1%) bekuyi-luminal A (Umfanekiso 3).Awekho amathumba e-HER2+ abekhona kuleli qembu.Ngakho, ukuguqulwa kwe-BRCA1 kuvame ezigulini ze-TNBC, kuyilapho ukuguqulwa kwe-BRCA2 kugqama kubantu abangabodwana be-lumen B.
Umfanekiso we-3 Ukuvama kwama-subtypes omdlavuza webele ezigulini ezinokuhlukahluka kwe-pathogenic ku-BRCA1 ne-BRCA2.I-Histograms ebonisa ukusabalalisa kwe-BRCA1- (obomvu obumnyama) kanye ne-BRCA2- (obomvu okhanyayo) ama-PVs phakathi kwama-molecular subtypes eziguli zomdlavuza webele.Izinombolo ezibikiwe ngaphakathi kwebhokisi ngalinye zimelela iphesenti leziguli ezine-BRCA1 subtype PV kanye ne-BRCA2 ngayinye yomdlavuza webele.
Izifinyezo: ama-PV, ukuhluka kwe-pathogenic;I-HER2+, i-epidermal growth factor receptor 2 positive;I-TNBC, umdlavuza webele ongenawo kathathu.
Ngokulandelayo, sihlole uhlobo nofuzo lwendawo lwe-BRCA1 kanye ne-BRCA2 PVs.Ku-BRCA1 PV, siqaphele okuhlukile okungu-7 kwe-nucleotide eyodwa (SNVs), ukususwa okungu-6, ukuphindaphinda okungu-3 kanye nokufakwa okungu-1. Ukuguqulwa okukodwa kuphela (c.5522delG) kumelela ukutholwa okusha okuvame ukutholwa530 kokubili kokutholwa okuvame kakhulu BRCA530 Kokubili i-P530 yesifundo. 9delCTAAT.Lokhu kuguqulwa kuhilela ukususwa kwama-nucleotide amahlanu (CTAAT) ku-BRCA1 exon 15, okuholela ekushintshweni kwe-amino acid leucine nge-tyrosine ku-codon 1679, futhi ngenxa yokuguqulwa kwefreyimu enenye indlela yokumisa ikhodoli ebikezelwe yokuholela ekuncipheni kwamaprotheni ngaphambi kwesikhathi, i-PtaV eyodwa kuphela ishintshile etholwe kutholwe enye iprotheni engaphambi kwesikhathi. isifunda sokuvumelana sesayithi ye-splice (c.4357+1G>T) (Ithebula 1).
Mayelana ne-BRCA2 PV, siqaphele ukususwa okungu-6, ama-SNV angu-6 nokuphindaphinda okungu-2. Azikho izinguquko ezitholakele eziyinoveli.Izinguquko ezintathu eziphinde zavela emphakathini wethu, c.428dup kanye no-c.8487+1G>A okuphawulwe ezihlokweni ezingu-3, ​​kulandelwa u-c.5851_5854ukushintshwa kwe-c. I-5 ye-BRCA2, ebikezelwe ukuthi izofaka ikhodi yephrotheni encishisiwe, engasebenzi.I-c.8487+1G>Ukuguqulwa kwenzeka endaweni ye-intronic ye-BRCA2 intron 19 (± 1,2) futhi kuthinta ukulandelana kwe-splicing ukuvumelana, okuholela ekuguquguqukeni kwe-splicing okuholela ku-absent545 ye-protein ye-absent515252525 ye-absentogenic ye-TTogenic. kuya ekususweni kwe-4-nucleotide kusuka ezikhundleni ze-nucleotide 5851 kuya ku-5854 ku-coding exon 10 yofuzo lwe-BRCA2 futhi iphumela ku-frameshift yokuhumusha nge-stop stop codon ebikezelwe (p.S1951WfsTer). Ngokuphawulekayo, njengoba kubikiwe ngaphambili, kokubili izinguquko> i-0 isiguli> i-C. Ukuguqulwa kokuqala kuhilela ukushintshwa kwe-adenosine (A) ku-BRCA2 exon 7 nge-guanine (G) equkethe i-nucleotide okuholela ekushintsheni kwe-valine ibe isoleucine ku-codon 211, i-isoleucine Amino acid i-amino acid enezici ezifanayo kakhulu.Lolu shintsho luthinta i-mRNA evamile ukuhlukana kwe-mRNA kanye nemiphumela ye-mRNA etholakala endaweni eyi-double substitution i-exstitution A yesibili (i-double substitute A) I-13 yofuzo efaka ikhodi ye-BRCA2.Ushintsho lwe-c.7008-2A>T lungase lukhiqize imibhalo eminingi yobude obuhlukahlukene.Ngaphezu kwalokho, eqenjini lama-BRCA2 PVs, izinguquko ezingu-4 kwezingu-18 (22.2%) beziyi-intronic.
Sibe sesenza imephu yokuguqulwa kwezinguquko ezisusayo ze-BRCA1/2 ezizindeni ezisebenzayo nasezifundeni ezibophezela amaprotheni (Fig. 4).Kufuzo lwe-BRCA1, ama-PV angu-50% abekwe endaweni yeqoqo lomdlavuza webele (BCCR), kuyilapho u-22% wokuguqulwa atholakala endaweni yeqoqo lomdlavuza we-ovarian (OCCR) (Fig. 32% ye-PCCR etholakala esifundeni se-BRCA52, i-PCR). kanye ne-42.8% yezinguquko zitholakala ku-OCCR (Fig. 4B).Okulandelayo, sihlole indawo ye-PV ngaphakathi kwesizinda samaprotheni e-BRCA1 kanye ne-BRCA2.Kuphrotheni ye-BRCA1, sithole ama-PV amathathu kusizinda se-loop kanye nekhoyili ehlanganisiwe, kanye nokuguqulwa okubili kusizinda se-BRCT (Umfanekiso, i-4A iphrotheni ye-BRCA iphinda i-4A). Izinguquko ze-intronic kanye ne-3 exonic zitholwe ku-oligo/oligosaccharide-binding (OB) kanye nezizinda zombhoshongo (T) (Figure 4B).
Umfanekiso 4 Ukumelwa okuhleliwe kwamaprotheni e-BRCA1 kanye ne-BRCA2 kanye nokwenza kwasendaweni kwezinhlobonhlobo ze-pathogenic.Lesi sibalo sibonisa ukusatshalaliswa kwe-BRCA1 (A) ne-BRCA2 (B) okuhlukile kwe-pathogenic ezigulini ezinomdlavuza webele.Ukuguqulwa kwe-Exonic kuboniswa ngombala oluhlaza okwesibhakabhaka, kuyilapho okuhlukile kwe-intronic kuboniswa ngowolintshi.Ubude bebha bumele inani lamacala.I-BRCA21 isizinda samaprotheni abikiwe futhi isizinda sawo se-BRCA21 esisebenzayo. isizinda se-loop (RING) kanye nokulandelana kwendawo yenuzi (i-NLS), isizinda sekhoyili ekhoyili, isizinda seqoqo le-SQ/TQ (SCD), kanye nesizinda se-BRCA1 C-terminal (BRCT).(B) Iphrotheni ye-BRCA2 iqukethe izimpinda eziyisishiyagalombili ze-BRC, isizinda esibophezela i-DNA esinesizinda se-helical (Helical), isizinda se-oligonding/OBoling-fold (i-OB) ne-toharide ephindwe kathathu I-NLS ohlangothini C.Izindawo ezibizwa ngokuthi i-Breast Cancer Cluster Region (BCCR) kanye ne-Ovarian Cancer Cluster Region (OCCR) zikhonjiswe ngezansi.*Imele ukuguqulwa kwezakhi zofuzo ezinquma ama-stop codon.
Sibe sesiphenya izici ze-BC clinicopathological ezingase zihlobane nokuba khona kwe-BRCA1/2 PV. Amarekhodi omtholampilo aphelele atholakalela iziguli eziyi-181 BRCA1/2-negative (ezingezona izithwali) kanye nabo bonke abathwali (n = 35).Kube khona ukuhlobana phakathi kwezinga lokwanda kwesimila kanye nebanga.
Sibale ukusatshalaliswa kwe-Ki-67 ngokusekelwe kumidiyeni yeqembu lethu (25%, ububanzi <10-90%).Izifundo ezine-Ki-67 <25% zichazwe ngokuthi “i-Ki-67″ ephansi, kuyilapho abantu abanamanani ≥ 25% babhekwa “njenge-Ki-67″ ephezulu.Umehluko Obalulekile0 (i-CAp-67) obalulekile (i-CAp-60) watholwa <i-1p-carriers engu-07 V07. abathwali (Fig. 5A).
Umfanekiso 5 Ukuhlobana kwe-Ki-67 nokusatshalaliswa kwebanga kwabesifazane abanomdlavuza webele abane-BRCA1 nangenayo i-BRCA2 PVs.(A) Ibhokisi elibonisa amanani amaphakathi e-Ki-67 ezigulini eziyi-181 ezingathwali ze-BC ngokumelene ne-BRCA1 (18) noma i-BRCA2 (17) iziguli ze-PV.P.P. iziguli ezinomdlavuza zibe ngamaqembu ebanga le-histological (G2 kanye ne-G3) ngokuya nge-BRCA1 kanye nesimo se-BRCA2 sokuguquka (izihloko ze-WT, i-BRCA1 kanye ne-BRCA2 PVs abathwali).
Ngokufanayo, sihlole ukuthi ingabe ibanga lesimila lihlobene nokuba khona kwe-BRCA1/2 PV.Njengoba i-G1 BC yayingekho emphakathini wethu, sihlukanise iziguli ngamaqembu amabili (G2 noma i-G3). Ngokuvumelana nemiphumela ye-Ki-67, ukuhlaziya kwembule ukuhlobana okubalulekile ngokwezibalo phakathi kwebanga lesimila kanye nokuguqulwa kwe-BRCA1, ngengxenye ephezulu ye-0rrierscarrier ye-01BR3 ye-G3 ye-G3. ) (Umfanekiso 5B).
Intuthuko kubuchwepheshe bokulandelana kwe-DNA yenze intuthuko engakaze ibonwe ekuhlolweni kofuzo kwe-BRCA1/2, okunomthelela obalulekile ezigulini ezinomlando womndeni womdlavuza. izifunda ezinemifanekiso.37 E-Italy, izinga le-BRCA1/2 PVs lalisuka ku-8% laya ku-37%, libonisa ukuhlukahluka okubanzi kwamazwe.38,39 Njengoba kunabantu abacishe babe izigidi ezingu-5, iSicily iyisifunda sesihlanu ngobukhulu e-Italy ngokwenani labantu abahlala khona.
Ucwaningo lwethu lungomunye wemibiko yokuqala mayelana nesigameko se-BRCA1/2 PV ezigulini ze-BC empumalanga yeSicily.28 Sigxile ekuhlaziyeni kwethu ku-BC, njengoba lokhu kuyisifo esivame kakhulu eqenjini lethu.
Lapho kuhlolwa iziguli ze-389 BC, i-9% iphethe i-BRCA1/2 PVs, isatshalaliswa ngokulinganayo phakathi kwe-BRCA1 ne-BRCA2.Le miphumela ihambisana naleyo ebibikwe ngaphambilini kubantu base-Italy.28 Kuyathakazelisa ukuthi, u-3% (13/389) weqembu lethu kwakungamadoda.Leli zinga liphakeme kunalokho okulindelekile kumdlavuza webele wabesilisa (1% 4 BRCA) ukukhethwa kwethu okusekelwe kumdlavuza webele we-BC (1% 4 BR0). Kodwa-ke, akekho kulawa madoda owakha i-BRCA1/2 PV, ngakho-ke ayengabakhandidethi bokuhlaziya okwengeziwe kwamangqamuzana ukuze kugwenywe ukuba khona kwezinguquko ezingavamile ezifana ne-PALB2, i-RAD51C kanye no-D, phakathi kwabanye.Izinhlobonhlobo zokubaluleka okungaqinisekile zitholwe ku-7% wezifundo lapho i-BRCA2 VUS yayibonakala khona.Ngisho nalo mphumela uhambisana28 nobufakazi obukhona,41.
Lapho sihlaziya ukusatshalaliswa kwe-BC molecular subtypes kwabesifazane abaguqukayo be-BRCA1/2, siqinisekise izinhlangano ezaziwayo phakathi kwe-TNBC ne-BRCA1 PV (58.8%) naphakathi kwe-luminal B BC kanye ne-BRCA2 PV (55.6%).16,43 I-luminal A ne-HER2+ izimila ku-BRCA1 kanye ne-BRCA1 ne-BRCA1 yenkampani yenethiwekhi 4 idatha ye-P2 yezincwadi ezikhona.
Bese sigxila ohlotsheni nendawo ye-BRCA1/2 PV.Eqenjini lethu, i-BRCA1 PV ejwayeleke kakhulu bekuyi-c.5035_5039delCTAAT.Nakuba u-Incorvaia et al.abazange bakuchaze lokhu okwehlukile eqenjini labo lase-Sicilian, abanye ababhali bakubike njengegciwane le-BRCA1 PV.34 Ama-PV amaningana e-BRCA1 atholwe eqenjini lethu - isb c.181T>G, c.514del, c.3253dupA kanye ne-c.5266dupC - okuye kwatholwa okubili kwe-1BRCA1 ku-Sici 28 (28) ku-Sicily. I-G kanye ne-c.5266dupC) ivame ukutholakala kumaJuda ase-Ashkenazi aseMpumalanga naseYurophu Ephakathi (iPoland, Czech), isiSlovenian, i-Austrian, isiHungary, iBelarusian kanye nesiJalimane ), i-44,45 futhi, e-United States nase-Argentina, isanda kuchazwa ngokuthi "i-germline variant" ezigulini zase-Italy ezine-BC ne-OC 34 iziguli ezinomdlavuza webele ngaphambili 34cant. ePalermo naseMessina.Ngokuthakazelisayo, ngisho no-Incorvaia et al.ithole okuhlukile kwe-c.3253dupA kweminye imindeni e-Catania.28 Ama-BRCA2 PV amelela kakhulu ngu-c.428dup, c.5851_5854delAGTT kanye nokwehluka oku-intronic c.8487+1G>A, okuye kwabikwa ngokuningiliziwe 28 esigulini e-Palerdup esine-c.528 yasendlini c.52V8 C. enyakatho-ntshonalanga yeSicily, ikakhulukazi ezifundeni zaseTrapani nasePalermo, kanti i-c.5851_5854delAGTT PV yabonwa emakhaya enyakatho-ntshonalanga yeSicily.Okuhlukile kwe-8487+1G>Akuvamile ezifundweni zaseMessina, Palermo, naseCaltanissetta.28 Rebbeck et al.ngaphambili kuchazwe ukuguqulwa kwe-c.5851_5854delAGTT e-Colombia.37 Enye i-BRCA2 PV, c.631+1G>A, itholwe ezigulini ze-BC kanye ne-OC ezivela eSicily (Agrigento, Siracusa and Ragusa).28 Ngokuphawulekayo, siye sabona ukuphilisana kokuhlukahluka kwe-BRCA2 kanye ne-31G0 yesiguli esifanayo (BRCA2>0 c.6>272) efanayo (BRCA2>0 efanayo). , ebesicabanga ukuthi sihlukaniswe ngemodi ye-cis, njengoba kubikwe kanjalo ngaphambili.34,46 Lezi zinguquko ze-BRCA2 uble zivame ukubonwa ngempela esifundeni sase-Italy futhi zitholwe ukuthi zethula ama-codons okuyeka ngaphambi kwesikhathi, okuthinta ukuhlanganiswa kwe-RNA yesithunywa futhi kubangela ukuthi iphrotheni ye-BRCA2 ihluleke.47,48
Siphinde futhi senze imephu ye-BRCA1 kanye ne-BRCA2 PVs ezindaweni ezibekayo ze-OCCR kanye ne-BCCR yezizinda zamaprotheni nezakhi zofuzo.Lezi zifunda zichazwe ngu-Rebbeck et al.njengezindawo eziyingozi zokuthuthukisa umdlavuza we-ovarian nowebele, ngokulandelana.49 Kodwa-ke, ubufakazi obuphathelene nokuhlangana phakathi kwendawo yokuhlukahluka kwegciwane kanye nengozi yomdlavuza webele noma we-ovarian kusalokhu kuyimpikiswano. Izifunda ze-BCCR nezici ze-BC.Lokhu kungase kube ngenxa yenani elilinganiselwe leziguli ezinokuguqulwa kwe-BRCA1/2. Ngokombono wesizinda samaprotheni, ama-BRCA1 PVs asatshalaliswa kuwo wonke amaprotheni, futhi ukuguqulwa kwe-BRCA2 kutholakala ngokukhethekile kusizinda sokuphinda se-BRC.
Ekugcineni, sihlobanise izici ze-BC clinicopathological ne-BRCA1/2 PV.Ngenxa yenani elilinganiselwe leziguli ezifakiwe, sithole kuphela ukuhlobana okubalulekile phakathi kwe-Ki-67 kanye nebanga lesimila. Nakuba ukuhlolwa nokuchazwa kwe-Ki-67 kusalokhu kuyimpikiswano ngandlela thile, kuqinisekile ukuthi amazinga aphezulu okukhula ahlotshaniswa nengozi eyandayo yokubuya kwesifo kanye nokwehla okukhulu "kokunciphisa ukubuya kwesifo". I-Ki-67 ingu-20%.Nokho, lo mkhawulo awusebenzi ku-BRCA1/2 yethu yesiguli esiguquguqukayo, enenani elimaphakathi le-Ki-67 elingu-25%.Lokhu kuthambekela kumazinga aphezulu we-Ki-67 kungachazwa ukwanda kweqembu lethu le-luminal B kanye ne-TNBC, lapho kwakukhona izimila ze-luminal A ezimbalwa. ekubikezelweni kwazo.53,54 Kusukela emiphumeleni yokuhlaziya kwethu, ukuhlobana okubalulekile akumangazi.Kuvela phakathi kwe-Ki-67 ephezulu kanye namamaki kanye nokuba khona kwe-BRCA1 PV.Eqinisweni, izimila ezihlobene ne-BRCA1 zivamile ku-TNBC futhi zibonisa izici ezinonya kakhulu.16,17
Sengiphetha, lolu cwaningo luhlinzeka ngombiko mayelana nesimo sokuguqulwa kwe-BRCA1/2 eqenjini le-BC kusukela empumalanga ye-Sicily. Sekukonke, okutholakele kwethu kuhambisana nobufakazi obukhona ngaphambili, kokubili mayelana nokusakazeka kokuguquka kanye nezici ze-clinicopathological ku-BC. Ucwaningo olwengeziwe kubantu abakhulu be-BRCA1 / 2-mutant-mutant BC, ukuhlaziya okuguquguqukayo kwe-PV okuguquguqukayo okuguquguqukayo okuguquguqukayo kweziguli eziguquguqukayo ze-BC, okufana nokuhlaziywa okuguquguqukayo kwe-PV okuguquguqukayo kweziguli eziningi ze-BC zihlukile futhi azivamile ukwedlula i-BRCA1/2.Lokhu kuzovumela ukuhlonza nokuphathwa ngendlela efanele kwenani elikhulayo lezifundo ezisengozini enkulu yomdlavuza ngenxa yokuguqulwa kofuzo.
Siqinisekise ukuthi iziguli zisayine imvume enolwazi ukuze zikhiphe amasampula azo isimila ngokungaziwa ngenjongo yocwaningo.Zonke iziguli zisayine imvume ebhaliwe enolwazi ngokweSimemezelo Sase-Helsinki.Ngokwenqubomgomo ye-AOU Policlinico "G.Rodolico - S.Marco", lolu cwaningo lukhishwe ekubuyekezweni kwezimiso zokuziphatha ngoba ukuhlaziya kwe-BRCA1/2 kwenziwa ngokuvumelana nezinjongo zocwaningo ezibhaliwe zeziguli futhi zanikeza imvume yocwaningo olubhaliwe lweziguli. .
Sibonga uProf. Paolo Vigneri ngosizo lwakhe ekunakekeleni iziguli ezinomdlavuza webele njengoba kucelwe iKomidi Lokuziphatha.
U-Federica Martorana ubika i-honoraria evela ku-Istituto Gentili, u-Eli Lilly, uNovartis, uPfizer.Abanye ababhali bamemezela ukuthi akukho ukungqubuzana kwezintshisekelo kulo msebenzi.
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