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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, 1 I-Medical Living Clinic1, 16 Umtholampilo we-Maperimental1 University of Catania, Catania, 95123, Italy;2 Centre for Experimental Oncology and Hematology, AOU Policlinico “G.Rodolico – San Marco”, Catania , 95123, Italy; 3 Medical Oncology, 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 Yezokwelapha Nezebhayoloji, iNyuvesi yaseCatania, Izakhi Zofuzo Zezokwelapha, iCatania, e-Italy, 95123; 7Oasi Research Institute-IRCCS, eTroina, 94018, e-Italy Ukuxhumana: Stefania Stella, ucingo +39 095 378 1946, i-imeyili [email protected]; [email protected] Inhloso: Ukuguqulwa kwezakhi zofuzo ku-BRCA1 kanye ne-BRCA2 kanye nokuthola umdlavuza webele (BC), i-ovary (OC) kanye nokunye okuhlobene nengozi yokuphila komdlavuza. Ukuhlolwa kwezakhi zofuzo ze-BRCA kubalulekile ekuhloleni ingozi yomuntu ngamunye, kanye nokuthola izindlela zokuvimbela kubantu abanempilo kanye nokwelashwa okwenziwa ngokwezifiso ezigulini zomdlavuza. Ukusabalala kokuguqulwa kwe-BRCA1 kanye ne-BRCA2 kuyahlukahluka kakhulu kuzo zonke izifunda, futhi yize idatha ikhona ngezinhlobo ze-BRCA ezibangela izifo emindenini yaseSicilian, izifundo eziqondiswe ngqo kubantu empumalanga yeSicily azitholakali. Inhloso yocwaningo lwethu kwakuwukuphenya ukwanda kanye nokusatshalaliswa kokuguqulwa kwezakhi zofuzo ze-BRCA ezibangela izifo eqenjini leziguli zase-BC ezivela empumalanga yeSicily nokuhlola ukuhlangana kwazo nezici ezithile ze-BC kusetshenziswa ukulandelana kwesizukulwane esilandelayo. Ukuba khona kokuguqulwa okuhlobene nebanga lesimila kanye nenkomba yokwanda. IMIPHUMELA: Sekukonke, iziguli ezingu-35 (9%) zazinohlobo lwe-BRCA olubangela izifo, ezingu-17 (49%) ku-BRCA1 kanye nezingu-18 (51%) ku-BRCA2. Ukuguqulwa kwe-BRCA1 kuvame kakhulu ezigulini ze-BC ezine-triple-negative, kanti ukuguquka kwe-BRCA2 kuvame kakhulu ezigulini ze-luminal BC. Uma kuqhathaniswa Kwabangebona abathwali, abantu abanezinhlobo ze-BRCA1 babenezinga eliphezulu kakhulu lesimila kanye nenkomba yokwanda kwaso. Iziphetho: Okutholakele kwethu kunikeza umbono jikelele wesimo sokuguquka kwe-BRCA ezigulini ze-BC ezivela empumalanga yeSicily futhi kuqinisekisa indima yokuhlaziywa kwe-NGS ekuboneni iziguli ezine-BC yofuzo. Sekukonke, le datha iyahambisana nobufakazi bangaphambilini obusekela ukuhlolwa kwe-BRCA ukuze kuvikelwe futhi kuphathwe kahle umdlavuza kubathwali bokuguquka kwezakhi zofuzo.
Umdlavuza webele (BC) umdlavuza ovame kakhulu emhlabeni wonke futhi uyingozi kakhulu kwabesifazane.1 Izici zebhayoloji ezinquma ukubikezela kwe-BC kanye nokuziphatha kwezokwelapha ziye zafundwa kabanzi futhi zacaciswa kancane ngokuhamba kwesikhathi.Eqinisweni, izimpawu eziningana ezisetshenziswayo njengamanje zisetshenziselwa ukuhlukanisa i-BC zibe izinhlobo ezahlukene zama-molecule.Ziyi-estrogen (ER) kanye/noma i-progesterone receptor (PgR), i-human epidermal growth factor receptor 2 (HER2) amplification, i-proliferation index Ki-67 kanye ne-tumor grade (G).2 Ukuhlanganiswa kwalezi ziguquguquko kubonise izigaba ezilandelayo ze-BC: 1) Izimila ze-Luminal, ezibonisa i-ER kanye/noma i-PgR expression, zabalelwa ku-75% wama-BC. Lezi zimila zahlukaniswa kabanzi zaba yi-Luminal A, lapho i-Ki-67 yayingaphansi kuka-20% kanye ne-HER2 negative, kanye ne-Luminal B, lapho i-Ki-67 yayilingana noma ingaphezu kuka-20% futhi lapho kukhona i-HER2 amplification, kungakhathaliseki i-proliferation index; 2) Izimila ze-HER2+ ezingenayo i-ER ne-PgR kodwa zibonisa ukwanda kwe-HER2. Leli qembu lihlanganisa u-10% wazo zonke izimila zebele; 3) Umdlavuza webele one-Triple-negative (TNBC), ongabonisi ukubonakaliswa kwe-ER ne-PgR kanye nokwanda kwe-HER2, uhlanganisa cishe u-15% womdlavuza webele.2-4
Phakathi kwalezi zinhlobo ze-BC, izinga lesimila kanye nenkomba yokwanda kumelela ama-biomarker anqamula isigaba ahlobene ngqo nangokuzimele nolaka lwesimila kanye nokubikezela.5,6
Ngaphezu kwezici zemvelo ezishiwo ngenhla, indima yokuguqulwa kwezakhi zofuzo okuholela ekuthuthukisweni kwe-BC iye yaba yinto ebaluleke kakhulu eminyakeni embalwa edlule.7 Cishe i-1 kweziyi-10 zezimila zesifuba zizuzwe njengefa ngenxa yokuguqulwa kwezakhi zofuzo kumajini athile.8 Izifundo ezimbili ezinkulu ze-epidemiological ezibandakanya abesifazane abangaphezu kuka-180,000 zisanda kukhomba iqembu lamajini ayisishiyagalombili (okungukuthi, i-ATM, i-BARD1, i-BRCA1, i-BRCA2, i-CHK2, i-PALB2, i-RAD51C, kanye ne-RAD51D) ezibangela kakhulu i-BC yofuzo. Phakathi kwalezi zakhi zofuzo, i-BRCA1 kanye ne-BRCA2 (ezobizwa ngokuthi i-BRCA1/2) zibonise ubudlelwano obuqine kakhulu nokuthuthukiswa kwezimila zesifuba.9-12 Eqinisweni, ukuguquka kwezakhi zofuzo kwe-BRCA1/2 kwandisa kakhulu ingozi yokuphila kwe-BC kanye nezinye izifo ezibangelwa umdlavuza, okuhlanganisa i-ovarian, i-prostate, i-pancreatic, i-colorectal, kanye ne-melanoma. Kusukela eminyakeni eyi-13 kuya kwengama-80, ukwanda kwe-BC kungu-72% kwabesifazane abane-BRCA1 pathogenic variant (PV) kanye no-69% kwabesifazane abane- I-BRCA2 PV.14
Okuphawulekayo ukuthi incwadi yakamuva isikisela ukuthi ingozi ye-BC incike ohlotsheni lwe-PV. Eqinisweni, uma kuqhathaniswa nezinhlobo ezinqamula izifo, izinhlobo ezingabonakali kahle, ikakhulukazi ku-gene ye-BRCA1, zihlotshaniswa nengozi encishisiwe ye-BC, ikakhulukazi kwabesifazane asebekhulile.15
Ukuba khona kwe-BRCA1 noma i-BRCA2 PV kuhlotshaniswa nezici ezahlukene zebhayoloji kanye nezokwelapha.16,17 Ama-BC ahlobene ne-BRCA1 avame ukuba nolaka ngokwezokwelapha, ahlukaniswe kahle, futhi anda kakhulu. Lawa mathumba avame ukuba ne-triple negative futhi aqala esemncane. Amathumba ayenzeka ezigulini eziguqukile ze-BRCA2 avame ukubonisa amamaki aphakathi kuya kwahluka kahle kanye nezinkomba zokwanda eziguquguqukayo. Lawa mathumba avame kakhulu ku-lumen B futhi avame ukwenzeka kubantu abadala.16-18 Okuphawulekayo ukuthi, ukuguqulwa kwezakhi zofuzo ku-BRCA1 naku-BRCA2 kwandisa ukuzwela ekwelashweni okuthile, kufaka phakathi usawoti we-platinum kanye nemithi eqondiwe efana ne-poly(ADP-ribose) polymerase inhibitors (PARPi).19,20
Eminyakeni embalwa edlule, ukusetshenziswa kwe-next-generation sequencing (NGS) emisebenzini yezokwelapha kuye kwenza inani elikhulayo leziguli ze-BC ukuthi lihlolwe ama-molecule ama-syndromes okuthi umuntu angakwazi ukutheleleka ngumdlavuza, okuhlanganisa i-BRCA1/2.21 Ngesikhathi esifanayo, izincazelo ezisekelwe ezindinganisweni eziqondile mayelana nomlando womndeni, izibalo zabantu, kanye nezici ze-clinicopathological ukuze kutholakale kangcono abantu abafanele ukuhlolwa kwe-BRCA1/2.22,23 Kulo mongo, ubufakazi buqongelela ekuhlolweni kwe-BRCA1/2 kubantu abathile, okugqamisa umehluko kuzo zonke izifunda zezwe.24–27 Nakuba kunemibiko ngeqembu le-BC entshonalanga yeSicily, kuncane idatha etholakalayo ekuhlolweni kwe-BRCA1/2 kubantu base-mpumalanga yeSicily.28,29
Lapha sichaza imiphumela yokuhlolwa kwe-germline BRCA1/2 ezigulini zase-BC ezivela empumalanga yeSicily, okuhlanganisa nokuba khona kwezinguquko ze-BRCA1 noma ze-BRCA2 nezici eziyinhloko zezokwelapha zalezi zimila.
Ucwaningo olubukezwa emuva lwenziwa e-“Center for Experimental Oncology and Hematology” ePoliclinico Hospital.Rodolico – San Marco eCatania. Kusukela ngoJanuwari 2017 kuya kuMashi 2021, iziguli ezingu-455 ezinesifo somdlavuza webele kanye ne-ovarian, i-melanoma, i-pancreatic noma i-prostate zathunyelwa elabhorethri yethu yokuxilonga ama-molecule ukuze zihlolwe izakhi zofuzo ze-BRCA/2. Lolu cwaningo lwenziwe ngokuhambisana neSimemezelo saseHelsinki, futhi bonke abahlanganyeli banikeze imvume ebhaliwe enolwazi ngaphambi kokuhlaziywa kwama-molecule.
Izici ze-histological kanye ne-biological (ER, PgR, isimo se-HER2, Ki-67, kanye nebanga) le-BC zihlolwe ku-biopsy eyinhloko noma amasampula okuhlinzwa, kucatshangelwa kuphela izingxenye zesimila esinamandla. Ngokusekelwe kulezi zici, ama-BC ahlukaniswe kanje: i-luminal A (ER+ kanye/noma i-PgR+, HER2-, Ki-67<20%), i-luminal B (ER+ kanye/noma i-PgR+, HER2-, Ki-67≥20%), i-luminal B-HER2+ (ER kanye/noma i-PgR+, HER2+), i-HER2+ (ER kanye ne-PgR-, HER2+) noma i-triple negative (ER kanye ne-PgR-, HER2-).
Ngaphambi kokuhlola isimo sokuguquka kwe-BRCA1 kanye ne-BRCA2, ithimba elihlanganisa isazi somdlavuza, isazi sezakhi zofuzo, kanye nodokotela wengqondo benze ukubonisana ngezakhi zofuzo zesimila kusiguli ngasinye ukuze kutholakale ukuthi kukhona i-BRCA1 kanye/noma i-BRCA1. noma abantu abanengozi ephezulu ye-PV ku-BRCA2 gene. Ukukhethwa kwesiguli kwenziwa ngokweziqondiso ze-Italian Society of Medical Oncology (AIOM) kanye nezincomo zendawo zaseSicilian.30,31 Lezi zindlela zifaka: (i) umlando womndeni wezinhlobo ezaziwayo zezifo ezibangelwa izifo kuma-genit susceptibility (isb., i-BRCA1, i-BRCA2, i-TP53, i-PTEN); (ii) abesilisa abane-BC; (iii) labo abane-BC kanye ne-OC; (iv) abesifazane abane-BC <36 iminyaka, i-TNBC <60 iminyaka, noma i-BC <50 iminyaka; (v) umlando wezokwelapha womuntu siqu we-BC <50 iminyaka kanye okungenani nesihlobo esisodwa sezinga lokuqala: (a) BC <50 iminyaka; (b) i-OC engekho mucinous futhi engekho emngceleni yanoma yimuphi ubudala; (c) i-BC yamazwe ngamazwe; (d) i-BC yamadoda; (e) umdlavuza we-pancreatic; (f) umdlavuza we-prostate; (vi) umlando womuntu siqu we-BC oneminyaka engaphezu kwengu-50 kanye nomlando womndeni we-BC, OC, noma umdlavuza we-pancreatic wezihlobo eziyizihlobo zezinga lokuqala komunye nomunye (kufaka phakathi izihlobo ahlobene nazo zezinga lokuqala); (vii) Umlando womuntu siqu we-OC kanye okungenani nesihlobo esisodwa sezinga lokuqala: (a) BC ngaphansi kweminyaka engu-50; (b) NOC; (c) i-BC yamazwe amabili; (d) i-BC yesilisa; (vii) owesifazane one-OC yezinga eliphezulu.
Isampula yegazi elingama-20 mL elivela esigulini ngasinye laqoqwa lafakwa emashubhu e-EDTA (BD Biosciences). I-Genomic DNA yahlukaniswa kumasampula egazi aphelele angu-0.7 mL kusetshenziswa i-QIAsymphony DSP DNA Midi kit Isolation Kit (QIAGEN, Hilden, Italy) ngokwemiyalelo yomenzi futhi yadlula nge-Qubit® 3.0 Fluorometer (Thermo Fisher Scientific, Waltham, MA, USA). Ukucebisa okuqondiwe kanye nokulungiselela umtapo wolwazi kwenziwa yi-Oncomine™ BRCA Research Assay Chef, elungele ukulayishwa kwi-Ion AmpliSeq™ Chef Reagents DL8 Kit ukuze kulungiselelwe umtapo wolwazi ngokuzenzakalelayo ngokwemiyalelo yomenzi. Le khithi iqukethe amachibi amabili e-multiplex PCR primer angasetshenziswa ukutadisha zonke izakhi zofuzo ze-BRCA1 (NM_007300.3) kanye ne-BRCA2 (NM_000059.3). Ngamafuphi, i-15 µL yesampula ngayinye ye-DNA ehlanjululwe (10 ng) yengezwe kumapuleti anebhakhodi ukuze kulungiselelwe umtapo wolwazi futhi wonke ama-reagents nezinto ezisetshenziswayo zalayishwa kuthuluzi le-Ion Chef™. Ukulungiswa komtapo wolwazi okuzenzakalelayo kanye nokuhlanganiswa kwesampula yomtapo wolwazi onebhakhodi kwenziwa kuthuluzi le-Ion Chef™. Inani lemitapo yolwazi elungisiwe labe selihlolwa yi-Qubit® 3.0 Fluorometer (Thermo Fisher Scientific, Waltham, MA, USA) ngokwemiyalelo yomenzi. Okokugcina, imitapo yolwazi ihlanganiswa ngezilinganiso ezilinganayo kumashubhu esampula omtapo wolwazi we-Ion Chef™. (amashubhu anebhakhodi) futhi alayishwa kuthuluzi le-Ion Chef™. Ukulandelana kwenziwa kusetshenziswa ithuluzi le-Ion Torrent S5 (Thermo Fisher Scientific) (Thermo Fisher Scientific) kusetshenziswa i-Ion 510 Chip (Thermo Fisher Scientific). Ukuhlaziywa kwedatha kwenziwe yi-Amplicon Suite (SmartSeq srl) kanye ne-Ion Reporter Software.
Zonke izinhlobo zokuqanjwa kwamagama zilandele iziqondiso zamanje ze-Human Genome Variation Consortium, etholakala ku-inthanethi (HGVS, http://www.hgvs.org/mutnomen). Ukubaluleka komtholampilo kwezinhlobo ze-BRCA1/2 kuchazwe kusetshenziswa ukuhlukaniswa kwe-International Consortium ENIGMA (Evidence-Based Network for Interpreting Germline Mutant Alleles, https://enigmaconsortium.org/) kanye nokubonisana ngemininingwane ehlukene efana ne-ARUP, BRCAEXCHANGE, ClinVar, IARC_LOVD, kanye ne-UMD. Ukuhlukaniswa kufaka phakathi izigaba ezinhlanu ezihlukile zengozi: i-benign (isigaba I), i-benign (isigaba II), i-variant of uncertainity significant (VUS, isigaba III), i-benign pathogenic (isigaba IV), kanye ne-pathogenic (isigaba V). I-VarAbanye baphinde bahlaziya umphumela wokuguqulwa kwezakhi zofuzo esakhiweni nasemsebenzini weprotheni, ithuluzi elinolwazi elinokufinyelela kumininingwane egciniwe engu-30.32
Ukuze kwabelwe ukubaluleka okungenzeka kwezokwelapha ku-VUS ngayinye, kusetshenziswe ama-algorithms okubikezela amaprotheni ekhompyutha alandelayo: I-MUTATION TASTER, 33 PROVEAN-SIFT (http://provean.jcvi.org/index.php), i-POLYPHEN-2 (http:// /genetics.bwh.harvard.edu/pph2/) kanye ne-Align-GVGD (http://agvgd.hci.utah.edu/agvgd_input.php). Izinhlobo ezihlukaniswe njengekilasi 1 nelesi-2 zazibhekwa njengezinhlobo zasendle.
Ukulandelana kweSanger kuqinisekisile ukuba khona kwe-pathogenic variant ngayinye. Ngamafuphi, kwaklanywa ama-primer amabili athile ku-variant ngayinye etholakele ngokusebenzisa i-BRCA1 kanye ne-BRCA2 gene reference sequences (NG_005905.2, NM_007294.3 kanye ne-NG_012772.3, NM_000059.3, ngokulandelana). Ngakho-ke, i-PCR eqondiwe yenziwa kwalandelwa i-Sanger sequencing.
Iziguli ezihlolwe zingenayo i-gene ye-BRCA1/2 zihlolwe nge-multiplex ligation-dependent probe amplification (MLPA) ngokwemiyalelo yomenzi ukuhlola ukuba khona kwe-large genomic rearrangements (LGR). Ngamafuphi, amasampula e-DNA ayasuswa futhi kusetshenziswa ama-probe aqondene ne-gene ye-BRCA1 ne-BRCA2 afinyelela ku-60, ngalinye lithola ukulandelana kwe-DNA ethile cishe ama-nucleotide angu-60 ubude. Imikhiqizo ye-probe amplification, equkethe isethi ehlukile yama-amplicons e-PCR, yabe isihlaziywa yi-capillary electrophoresis kanye nesofthiwe ye-Cofalyser.Net ngokubambisana namathebula e-Cofalyser afanele (www.mrcholland.com).
Iziguquguquko ze-clinicopathological ezikhethiwe (ibanga le-histological kanye ne-Ki-67% proliferation index) zihlotshaniswa nokuba khona kwe-BRCA1/2 PV, kubalwa kusetshenziswa isofthiwe ye-Prism v. 8.4 kusetshenziswa ukuhlolwa okuqondile kukaFisher okuthatha inani le-p <0.05 njengelibalulekile.
Phakathi kukaJanuwari 2017 noMashi 2021, iziguli ezingu-455 zahlolwa i-germline BRCA1/2 mutations. Ukuhlolwa kokuguqulwa kwezakhi zofuzo kwenziwa ePoliclinico Hospital's Center for Experimental Oncology and Hematology. Ngokusho kwesiqondiso saseSicilian (http://www.gurs.regione.sicilia.it/Indicep1.htm, N. 02-Venerdì 10 Gennaio 2020), iRodolico yaseCatania – San Marco” iyonke, iziguli ezingu-389 Kwakukhona umdlavuza webele, umdlavuza wesibeletho ongu-37, umdlavuza we-pancreatic ongu-16, umdlavuza we-prostate ongu-8 kanye ne-melanoma engu-5. Ukusatshalaliswa kweziguli ngokohlobo lomdlavuza kanye nemiphumela yokuhlaziya kuboniswe kuMfanekiso 1.
Isibalo 1 sibonisa ishadi lokugeleza elibonisa ukubuka konke kwalolu cwaningo. Iziguli ezinezimila zebele, i-melanoma, i-pancreatic, i-prostate, noma ama-ovarian zihlolwe izinguquko ezakhiweni ze-BRCA1 kanye ne-BRCA2.
Izifinyezo: ama-PV, uhlobo olubangela izifo; i-VUS, uhlobo olungenaso isiqiniseko; i-WT, uhlobo lwe-wild-type BRCA1/2 sequence.
Sigxile ngokukhetha izifundo zethu kumaqoqo omdlavuza webele. Iziguli zazineminyaka ephakathi kwengu-49 (ibanga eliphakathi kuka-23-89) futhi iningi lazo kwakungabesifazane (n=376, noma 97%).
Kulaba bantu, abangu-64 (17%) babenezinguquko ze-BRCA1/2 futhi bonke babengabesifazane. Abangamashumi amathathu nanhlanu (9%) babene-PV kanti abangu-29 (7.5%) babene-VUS. Ishumi nesikhombisa (48.6%) kwezinhlobo ezingu-35 ze-pathogenic zenzeka ku-BRCA1 kanye no-18 (51.4%) ku-BRCA2, kanti abangu-5 be-VUS benzeka ku-BRCA1 (17.2%) kanye no-24 (82.8%) ku-BRCA2 (Izibalo 1 no-2). I-LGR yayingekho ekuhlaziyweni kwe-MLPA.
Umfanekiso 2. Ukuhlaziywa kwezinguquko ze-BRCA1 kanye ne-BRCA2 ezigulini ezingu-389 zomdlavuza webele.(A) Ukusatshalaliswa kwezinhlobo ze-pathogenic (PV) (obomvu), izinhlobo ezingaqinisekile (VUS) (ophuzi), kanye ne-WT (oluhlaza okwesibhakabhaka) ezigulini ezingu-389 zomdlavuza webele; (B) Iziguli ezingu-389 zomdlavuza webele Ama-35 (9%) ayenezinhlobo ze-BRCA1/2 ze-pathogenic (ama-PV).Phakathi kwazo, ezingu-17 (48.6%) zaziyi-BRCA1 PV carriers (obomvu obumnyama) kanti ezingu-18 (51.4%) zaziyi-BRCA2 carriers (obomvu okhanyayo); (C) Abantu abangu-29 (7.5%) kwabangu-389 ababene-VUS, izakhi zofuzo ze-BRCA1 ezingu-5 (17.2%) (obomvu obumnyama) kanye nezakhi zofuzo ze-BRCA2 ezingu-24 (82.8%) (obomvu okhanyayo).
Izifinyezo: ama-PV, uhlobo olubangela izifo; i-VUS, uhlobo olungenaso isiqiniseko; i-WT, uhlobo lwe-wild-type BRCA1/2 sequence.
Ngokulandelayo sihlole ukusabalala kwama-subtypes ama-molecular e-BC ezigulini ezine-BRCA1/2 PV. Ukusatshalaliswa kwakuhlanganisa iziguli ezimbili (5.7%) ze-luminal A, eziyi-15 (42.9%) ze-luminal B, eziyi-3 (8.6%) ze-luminal B-HER2+, eziyi-2 (5.7%) ze-HER2+ kanye neziyi-13 (37.1%) ze-TNBC. Phakathi kweziguli ezine-BRCA1, eziyi-5 (29.4%) zazine-luminal B BC, eziyi-2 (11.8%) zazinesifo se-HER2+, kanti eziyi-10 (58.8%) zazine-TNBC. Ama-tumors angenayo i-BRCA1 mutations kwakuyi-luminal A noma i-luminal B-HER2+ (Isithombe 3). Eqenjini elincane le-BRCA2-positive, izimila eziyi-10 (55.6%) zaziyi-luminal B, eziyi-3 (16.7%) zaziyi-luminal B-HER2+, eziyi-3 (16.7%) ze-TNBC kanye nezi-2 (11.1%) zaziyi-luminal A (Isithombe 3). Cha. Izimila ze-HER2+ zazikhona kuleli qembu. Ngakho-ke, izinguquko ze-BRCA1 zivame kakhulu ezigulini ze-TNBC, kanti izinguquko ze-BRCA2 zivame kakhulu kubantu abane-lumen B.
Isibalo 3 Ukusabalala kwezinhlobo zomdlavuza webele ezigulini ezinezinhlobo ezibangela izifo ku-BRCA1 kanye ne-BRCA2. Ama-histogram abonisa ukusatshalaliswa kwe-BRCA1- (ubomvu obumnyama) kanye ne-BRCA2- (ubomvu obukhanyayo) PV phakathi kwezinhlobo zama-molecule zeziguli zomdlavuza webele. Izinombolo ezibikiwe ngaphakathi kwebhokisi ngalinye zimele iphesenti leziguli ezine-BRCA1 kanye ne-BRCA2 PV yohlobo ngalunye lomdlavuza webele.
Izifinyezo: Ama-PV, uhlobo lwe-pathogenic; i-HER2+, i-human epidermal growth factor receptor 2 positive; i-TNBC, umdlavuza webele one-triple-negative.
Ngemva kwalokho, sihlole uhlobo kanye nendawo yezakhi zofuzo ze-BRCA1 kanye ne-BRCA2 PV. Ku-BRCA1 PV, sibone izinhlobo ezingu-7 ze-nucleotide eyodwa (ama-SNV), ukususwa okungu-6, ukuphindaphindwa okungu-3 kanye nokufakwa okukodwa. Ukuguqulwa okukodwa kuphela (c.5522delG) kumelela ukutholakala okusha. I-BRCA1 PV evame kakhulu etholakale kuzo zombili izifundo kwakuyi-c.5035_5039delCTAAT. Lolu shintsho luhilela ukususwa kwama-nucleotide amahlanu (i-CTAAT) ku-BRCA1 exon 15, okwaholela ekufakweni kwe-amino acid leucine yi-tyrosine ku-codon 1679, futhi ngenxa yokushintsha kohlaka lokuhumusha nge-alternative stop codon okubikezelwe kuholela ekunqunyweni kwamaprotheni ngaphambi kwesikhathi. Zonke ezinye izinguquko zitholakala esimweni esisodwa kuphela. Okuphawulekayo ukuthi enye yama-PV abikiwe yayitholakala esifundeni se-splice site consensus (c.4357+1G>T) (Ithebula 1).
Ngokuphathelene ne-BRCA2 PV, sibone ukususwa okungu-6, ama-SNV angu-6 kanye nokuphindaphindwa okungu-2. Akukho noyedwa kulezo zinguquko ezitholakele okwethusha. Ukuguqulwa okuthathu kuphinde kwenzeka kubantu bethu, i-c.428dup kanye ne-c.8487+1G>A ebonwe kubantu abangu-3, ​​kulandelwa yi-c.5851_5854delAGTT etholakale ezimweni ezimbili. Ukuguqulwa kwe-c.428dup kuhilela ukuphindaphindwa kwe-C ku-exon 5 ye-BRCA2, okubikezelwe ukuthi izofaka ikhodi yephrotheni enqunyiwe, engasebenzi. Ukuguqulwa kwe-c.8487+1G>A kwenzeka esifundeni se-intronic se-BRCA2 intron 19 (± 1,2) futhi kuthinta ukulandelana kokuvumelana kwe-splicing, okuholela ekushintsheni kwe-splicing okuholela kuphrotheni engavamile noma engekho. Uhlobo lwe-c.5851_5854delAGTT oluyimbangela yokususwa kwe-nucleotide engu-4 ezindaweni ze-nucleotide ezingu-5851 kuya ku-5854 ku-exon 10 yekhodi I-gene ye-BRCA2 futhi iphumela ekushintsheni kohlaka lokuhumusha nge-codon yokuma ehlukile ebikezelwe (p.S1951WfsTer). Okuphawulekayo, njengoba kubikiwe ngaphambilini, zombili izinguquko c.631G>A kanye c.7008-2A>T zitholakale esigulini esifanayo.34 Uguquko lokuqala luhilela ukufakwa esikhundleni kwe-adenosine (A) ku-exon 7 ye-BRCA2 nge-guanine (G) equkethe i-nucleotide okuholela ekushintsheni kwe-valine ibe yi-isoleucine ku-codon 211, i-isoleucine I-Amino acid iyi-amino acid enezakhiwo ezifanayo kakhulu. Lolu shintsho luthinta ukuhlanganiswa okuvamile kwe-mRNA. Uhlobo lwesibili lutholakala esifundeni se-intronic futhi luholela ekushintsheni kwe-A kabili ku-thymine (T) ngaphambi kwe-exon 13 ye-gene efaka i-BRCA2. Ushintsho lwe-c.7008-2A>T lungakhiqiza ama-transcript amaningi anobude obuhlukene. Ngaphezu kwalokho, eqenjini lama-PV e-BRCA2, izinguquko ezi-4 kweziyi-18 (22.2%) zaziyi-intronic.
Sabe sesidweba i-BRCA1/2 defectious mutations ezindaweni ezisebenzayo kanye nasezindaweni ezibopha amaprotheni (Isithombe 4). Ku-BRCA1 gene, ama-50% ama-PV atholakala esifundeni seqembu lomdlavuza webele (i-BCCR), kanti ama-22% ama-mutation atholakala esifundeni seqembu lomdlavuza we-ovarian (i-OCCR) (Isithombe 4A). Ku-BRCA2 PV, ama-35.7% ezinhlobonhlobo atholakala esifundeni se-BCCR kanti ama-42.8% ama-mutation atholakala ku-OCCR (Isithombe 4B). Okulandelayo, sihlole indawo ye-PV ngaphakathi kwezizinda zephrotheni ye-BRCA1 kanye ne-BRCA2. Kuphrotheni ye-BRCA1, sithole ama-PV amathathu ezizindeni ze-loop kanye ne-coiled coil, kanye nezinguquko ezimbili esizindeni se-BRCT (Isithombe 4A). Kuphrotheni ye-BRCA2, ama-PV amane afakwe ku-BRC repeat domain, kanti izinguquko ezintathu ze-intronic kanye nezingu-3 ze-exonic zitholakale ezizindeni ze-oligo/oligosaccharide-binding (OB) kanye ne-tower (T) (Isithombe 4B).
Isithombe 4 Ukumelwa kwe-schematic kwamaprotheni e-BRCA1 kanye ne-BRCA2 kanye nokwenziwa kwendawo kwezinhlobo ze-pathogenic. Lesi sibalo sibonisa ukusatshalaliswa kwezinhlobo ze-BRCA1 (A) kanye ne-BRCA2 (B) ezifweni zomdlavuza webele. Ukuguqulwa kwe-exonic kuboniswa ngombala oluhlaza okwesibhakabhaka, kuyilapho izinhlobo ze-intronic kuboniswa ngombala ophuzi. Ukuphakama kwebha kumelela inani lamacala. Amaprotheni e-BRCA1 kanye ne-BRCA2 kanye nezizinda zawo ezisebenzayo kubikiwe. (A) Iphrotheni ye-BRCA1 iqukethe isizinda se-loop (RING) kanye nochungechunge lwendawo ye-nuclear (NLS), isizinda se-coiled-coil, isizinda se-SQ/TQ cluster (SCD), kanye nesizinda se-BRCA1 C-terminal (BRCT). (B) Iphrotheni ye-BRCA2 iqukethe ukuphindaphinda kwe-BRC okuyisishiyagalombili, isizinda esibopha i-DNA esinesizinda se-helical (Helical), ama-fold amathathu e-oligonucleotide/oligosaccharide-binding (OB), isizinda se-tower (T), kanye ne-An NLS ohlangothini lwe-C. Izindawo ezibizwa ngokuthi i-Breast Cancer Cluster Region (BCCR) kanye ne-Ovarian Cancer Cluster Region (OCCR) ziboniswe ku- phansi.* Kumelela izinguquko ezinquma ama-stop codon.
Sabe sesihlola izici ze-BC clinicopathological ezingase zihlobane nokuba khona kwe-BRCA1/2 PV. Amarekhodi aphelele emitholampilo ayetholakala ezigulini ezingu-181 ezingenayo i-BRCA1/2 (ezingenazo izithwali) kanye nabo bonke abathwali (n = 35). Kwakukhona ubudlelwano phakathi kwesilinganiso sokukhula kwesimila kanye nebanga.
Sibale ukusatshalaliswa kwe-Ki-67 ngokusekelwe ku-median yeqembu lethu (25%, ububanzi <10-90%). Izihloko ezine-Ki-67 <25% zichazwe njenge-"Ki-67 ephansi", kuyilapho abantu abanamanani angu-≥ 25% babhekwa njenge-"Ki-67 ephezulu". Umehluko omkhulu we-Ki-67 (p<0.01) utholakale phakathi kwabangebona abathwali kanye nabathwali be-BRCA1 PV (Isithombe 5A).
Isithombe 5 Ubudlelwano be-Ki-67 nokusatshalaliswa kwebanga kwabesifazane abanomdlavuza webele abane-BRCA1 kanye ne-BRCA2 PV nabangenayo.(A) Ibhokisi elibonisa amanani aphakathi kwe-Ki-67 ezigulini ezingu-181 ze-BC ezingabathwali uma kuqhathaniswa neziguli ze-BRCA1 (18) noma ze-BRCA2 (17) ze-PV. Amanani e-P angaphansi kuka-0.5 abhekwa njengabalulekile ngokwezibalo.(B) I-Histogram emele ukwabiwa kweziguli zomdlavuza we-BC emaqenjini ebanga le-histological (G2 kanye ne-G3) ngokusho kwesimo sokuguquka kwe-BRCA1 kanye ne-BRCA2 (izihloko ze-WT, abathwali be-BRCA1 kanye ne-BRCA2 PVs).
Ngokufanayo, sihlole ukuthi izinga lesimila liyahambisana yini nokuba khona kwe-BRCA1/2 PV. Njengoba i-G1 BC yayingekho kubantu bethu, sahlukanisa iziguli ngamaqembu amabili (i-G2 noma i-G3). Ngokuhambisana nemiphumela ye-Ki-67, ukuhlaziywa kwembule ubudlelwano obubalulekile ngokwezibalo phakathi kwebanga lesimila kanye nokuguqulwa kwe-BRCA1, kanye nengxenye ephezulu yezimila ze-G3 kubathwali be-BRCA1 uma kuqhathaniswa nabangebona abathwali (p<0.005) (Isithombe 5B).
Intuthuko kwezobuchwepheshe bokulandelana kwe-DNA ivumele intuthuko engakaze ibonwe ekuhlolweni kwezakhi zofuzo ze-BRCA1/2, kanye nemiphumela ebalulekile ezigulini ezinomlando womndeni womdlavuza. Kuze kube manje, cishe izinhlobo ze-BRCA1/2 ezingaba ngu-20,000 zitholakale futhi zahlukaniswa ngokwe-American Society of Medical Genetics 35 kanye nohlelo lwe-ENIGMA.35,36 Kwaziwa kahle ukuthi i-BRCA1/2 mutational spectrum iyahlukahluka kakhulu kuzo zonke izifunda zezwe.37 Ngaphakathi kwe-Italy, izinga lama-PV e-BRCA1/2 lalisukela ku-8% kuya ku-37%, okubonisa ukuhlukahluka okukhulu kwangaphakathi kwezwe.38,39 Njengoba inabantu abacishe babe yizigidi ezi-5, iSicily iyisifunda sesihlanu ngobukhulu e-Italy ngokwenani labantu abahlala khona. Nakuba kukhona idatha ngokusatshalaliswa kwe-BRCA1/2 entshonalanga yeSicily, abukho ubufakazi obuningi engxenyeni esempumalanga yesiqhingi.
Ucwaningo lwethu lungomunye wemibiko yokuqala mayelana nokwanda kwe-BRCA1/2 PV ezigulini ze-BC empumalanga yeSicily.28 Sigxile ekuhlaziyeni kwethu ku-BC, njengoba lesi yisifo esivame kakhulu eqenjini lethu.
Lapho kuhlolwa iziguli ezingu-389 BC, u-9% wayephethe i-BRCA1/2 PV, esatshalaliswe ngokulinganayo phakathi kwe-BRCA1 ne-BRCA2. Le miphumela ihambisana naleyo ebikwe ngaphambilini kubantu base-Italy.28 Ngokuthakazelisayo, u-3% (13/389) weqembu lethu babengabesilisa. Leli zinga liphakeme kunalokho obekulindelwe kumdlavuza webele wabesilisa (1% wabo bonke ama-BC),40 okubonisa ukukhetha kwethu kwabantu ngokusekelwe engcupheni yokuguquka kwe-BRCA1/2. Kodwa-ke, akekho kulaba besilisa owasungula i-BRCA1/2 PV, ngakho-ke babengabalingani bokuhlaziywa okwengeziwe kwama-molecule ukuze kunqunywe ukuba khona kokuguquka kwezakhi zofuzo okungajwayelekile njenge-PALB2, RAD51C kanye ne-D, phakathi kwabanye. Izinhlobo ezingaqinisekile zitholwe ku-7% wezifundo lapho i-BRCA2 VUS yayibonakala khona. Ngisho nalo mphumela uhambisana nobufakazi obukhona.28,41,42
Lapho sihlaziya ukusatshalaliswa kwama-subtypes e-BC molecular kwabesifazane abaguqukile be-BRCA1/2, siqinisekisile ubudlelwano obaziwayo phakathi kwe-TNBC ne-BRCA1 PV (58.8%) kanye naphakathi kwe-luminal B BC ne-BRCA2 PV (55.6%).16,43 Ama-luminal A kanye ne-HER2+ amathumba ku-BRCA1 kanye ne-BRCA2 PV carriers ahambisana nedatha ekhona yezincwadi.16,43
Sibe sesigxila ohlotsheni nasendaweni ye-BRCA1/2 PV. Eqenjini lethu, i-BRCA1 PV evame kakhulu kwakuyi-c.5035_5039delCTAAT. Nakuba u-Incorvaia et al. abazange bachaze lolu hlobo lwe-germline eqenjini labo laseSicilian, abanye ababhali babike ukuthi luyi-germline BRCA1 PV.34 Kutholakale ama-PV amaningana e-BRCA1 eqenjini lethu – isib. c.181T>G, c.514del, c.3253dupA kanye ne-c.5266dupC – okuye kwabonwa eSicily.28 Kulawa, izinguquko ezimbili zabasunguli be-BRCA1 (c.181T>G kanye ne-c.5266dupC) zivame ukutholakala kumaJuda ase-Ashkenazi aseMpumalanga nase-Europe Ephakathi (ePoland, eCzech), eSlovenia, e-Austria, eHungary, eBelarusian naseJalimane), 44,45 futhi, e-United States nase-Argentina, muva nje kuchazwe ngokuthi “uhlobo lwe-germline oluphindaphindayo” ezigulini zase-Italy ezine-BC kanye ne-OC. Uhlobo lwe-34c.514del lwatholakala ngaphambilini ezigulini ezingu-8 zomdlavuza webele ezivela enyakatho yeSicily ePalermo naseMessina. Kuyathakazelisa ukuthi ngisho no-Incorvaia et al. bathole uhlobo lwe-c.3253dupA kweminye imindeni eCatania.28 Ama-PV e-BRCA2 amele kakhulu yi-c.428dup, c.5851_5854delAGTT kanye nohlobo lwe-intronic c.8487+1G>A, olubikwe ngokuningiliziwe 28 esigulini ePalermo esine-c.428dup, c.5851_5854delAGTT PV yabonwa emizini esenyakatho-ntshonalanga yeSicily, ikakhulukazi ezifundeni zaseTrapani nasePalermo, kanti i-c.5851_5854delAGTT PV yabonwa emizini esenyakatho-ntshonalanga yeSicily. Uhlobo lwe-8487+1G>A lwaluvame kakhulu ezifundweni ezivela eMessina, ePalermo, naseCaltanissetta.28 Rebbeck et al. ngaphambilini kuchazwe ukuguqulwa kwe-c.5851_5854delAGTT eColombia.37 Enye i-BRCA2 PV, c.631+1G>A, itholakale ezigulini ze-BC kanye ne-OC ezivela eSicily (Agrigento, Siracusa kanye neRagusa).28 Okuphawulekayo ukuthi sibone ukuba khona kwezinhlobo ezimbili ze-BRCA2 (BRCA2 c.631G>A kanye c.7008-2A>T) esigulini esifanayo, esasicabanga ukuthi sihlukaniswe kwimodi ye-cis, njengoba kubikwe ngaphambili kanjalo.34,46 Lezi zinguquko ze-BRCA2 zivame ukubonwa esifundeni sase-Italy futhi zitholakale ukuthi zethula ama-codon okuma ngaphambi kwesikhathi, zithinta ukuxhunyaniswa kwe-messenger RNA futhi zibangele ukuthi iphrotheni ye-BRCA2 yehluleke.47,48
Siphinde sahlela ama-PV e-BRCA1 kanye ne-BRCA2 ezindaweni ze-OCCR kanye ne-BCCR ezifakazelwe yizizinda zamaprotheni nezakhi zofuzo. Lezi zindawo zachazwa nguRebbeck et al. njengezindawo eziyingozi zokuthuthukisa umdlavuza we-ovarian kanye nebele, ngokulandelana.49 Kodwa-ke, ubufakazi obuphathelene nobudlelwano phakathi kwendawo yezinhlobo ze-germline kanye nengozi yomdlavuza webele noma we-ovarian busalokhu buphikisana.28,50-52 Emphakathini wethu, ama-PV e-BRCA1 ayetholakala kakhulu esifundeni se-BCCR, kanti ama-PV e-BRCA2 ayetholakala kakhulu esifundeni se-OCCR. Kodwa-ke, asikwazanga ukuthola noma yikuphi ubudlelwano phakathi kwezifunda ze-OCCR kanye ne-BCCR ezifakazelwe kanye nezici ze-BC. Lokhu kungenzeka ngenxa yenani elilinganiselwe leziguli ezinezinguquko ze-BRCA1/2. Kusukela embonweni wesizinda seprotheni, ama-PV e-BRCA1 asatshalaliswa kulo lonke iphrotheni, kanti izinguquko ze-BRCA2 zitholakala ngokukhethekile endaweni yokuphindaphinda ye-BRC.
Ekugcineni, sihlanganise izici ze-BC clinicopathological ne-BRCA1/2 PV. Ngenxa yenani elilinganiselwe leziguli ezifakiwe, sithole kuphela ubudlelwano obubalulekile phakathi kwe-Ki-67 kanye nezinga lesimila. Nakuba ukuhlolwa nokuchazwa kwe-Ki-67 kusalokhu kuyimpikiswano ethile, kuqinisekile ukuthi amazinga aphezulu okwanda ahlotshaniswa nengozi eyengeziwe yokuphinda kwesifo kanye nokuncipha kokusinda. Kuze kube manje, ukuphela kokuhlukanisa phakathi kwe-Ki-67 "ephezulu" kanye "nephansi" kungu-20%. Kodwa-ke, lo mkhawulo awusebenzi kubantu bethu be-BRCA1/2 mutation, abanenani eliphakathi le-Ki-67 elingu-25%. Lo mkhuba wamazinga aphezulu e-Ki-67 ungachazwa ngokusabalala kwamaqembu ethu e-luminal B kanye ne-TNBC, lapho kwakukhona khona izimila ezimbalwa ze-luminal A. Kodwa-ke, ubufakazi obuthile bubonakala busikisela ukuthi ukuhlukaniswa okuphezulu kwe-Ki-67 (25-30%) kungahlukanisa kangcono iziguli ngokwesimo sazo.53,54 Kusukela emiphumeleni yokuhlaziywa kwethu, ubudlelwano obubalulekile abumangazi. Kwenzeka phakathi kwe-Ki-67 ephezulu kanye namabanga kanye nokuba khona kwe-BRCA1 I-PV.Eqinisweni, izimila ezihlobene ne-BRCA1 zivamile kwi-TNBC futhi zibonisa izici ezinolaka kakhulu.16,17
Ekuphetheni, lolu cwaningo luhlinzeka ngombiko ngesimo sokuguquka kwe-BRCA1/2 eqenjini le-BC elivela empumalanga yeSicily. Sekukonke, okutholakele kwethu kuhambisana nobufakazi obukhona, kokubili maqondana nokusabalala kokuguquka kwezakhi zofuzo kanye nezici ze-clinicopathological e-BC. Izifundo ezengeziwe kubantu abaningi beziguli ze-BRCA1/2-mutant BC, njengokusebenzisa ukuhlaziywa kokuguquguquka kwezakhi zofuzo okunwetshiwe okuningi, ziyafuneka ukuhlola ukuba khona kwama-PV ahlukile futhi angavami kakhulu kune-BRCA1/2. Lokhu kuzovumela ukuhlonza nokuphathwa okufanele kwenani elikhulayo labantu abasengozini enkulu yomdlavuza ngenxa yokuguquka kwezakhi zofuzo.
Siqinisekisile ukuthi iziguli zisayine imvume enolwazi yokukhipha amasampula azo esimila ngokungaziwa ngezinjongo zocwaningo. Zonke iziguli zisayine imvume ebhaliwe enolwazi ngokweSimemezelo saseHelsinki. Ngokwenqubomgomo ye-AOU Policlinico “G.Rodolico – S.Marco”, lolu cwaningo aluzange luvunyelwe ukubuyekezwa kokuziphatha ngoba ukuhlaziywa kwe-BRCA1/2 kwenziwa ngokwemikhuba yezokwelapha futhi zonke iziguli zinikeze imvume ebhaliwe enolwazi. Iziguli nazo ziyavuma ukusetshenziswa kwedatha yazo ngezinjongo zocwaningo.
Sibonga uSolwazi Paolo Vigneri ngosizo lwakhe ekunakekelweni kweziguli ezinomdlavuza webele njengoba kuceliwe yiKomidi Lezokuziphatha.
UFederica Martorana ubika nge-honaria evela ku-Istituto Gentili, u-Eli Lilly, uNovartis, uPfizer. Abanye ababhali bathi akukho ukungqubuzana kwezintshisekelo kulo msebenzi.
1. Sung H, Ferlay J, Siegel RL, nabanye. Izibalo Zomdlavuza Womhlaba Wonke 2020: I-GLOBOCAN ilinganisela ukwenzeka kanye nokufa kwezinhlobo zomdlavuza ezingu-36 emazweni angu-185 emhlabeni jikelele. I-CA Cancer J Clin.2021;71(3):209-249.doi: 10.3322/caac.21660