▎ PNC 27 Okunoonyereza
PNC-27 ekozesebwa ki?
1, Obujjanjabi bw’obulwadde bwa leukemia:
PNC-27 esobola okusiba puloteyina ya HDM-2 ku luwuzi lw’obutoffaali bwa kookolo n’okuleeta okutondebwa kw’obutuli obuyita mu lususu obuyitibwa cytotoxic transmembrane pores. Mu kunoonyereza okwakolebwa ku layini z’obutoffaali bw’omuntu obutali bwa stem cell acute myeloid leukemia, HDM-2 yazuulibwa nga yeeyolekera nnyo mu membranes za U937 (acute monocytic leukemia), OCI-AML3 (acute granulomonocytic leukemia), ne HL60 (acute promyelocytic leukemia) cells.PNC-27 esiba ku HDM-2 ku membranes, era n’ereetera necroptosis ne LDH (lactate dehydrogenase) okufuluma mu ssaawa 4. (LDH) okufulumya mu ssaawa 4. Okutunuulira HDM-2 ku luwuzi lw’obutoffaali kiyinza okuba akakodyo akayinza okujjanjaba obulwadde bwa leukemia.PNC-27 yalaga omulimu omukulu ogw’okulwanyisa obulwadde bwa leukemia mu layini z’obutoffaali bwa leukemia eziwerako nga etunuulira Membrane HDM-2 [1]..
2. Kireetera obutoffaali bw’ekizimba okusasika
Akola nga peptide etaliiko kamogo:
Okunoonyereza okuwerako kulaga nti PNC-27 ereeta okukutuka kw’obutoffaali bw’ekizimba nga peptide etaliiko kamogo okusinga ng’akatundu. Okugeza, mu kunoonyereza ku butoffaali bwa kookolo w’amabeere MCF-7 n’obutoffaali bw’amabeere obutakyusiddwa MCF-10-2A, obutoffaali bwajjanjabwa ne PNC-27 nga bulina amino terminus eya green fluorescently labeled ne red fluorescently labeled carboxy terminus. Ebivuddemu biraga ekitangaala eky’enjawulo ekya kiragala ekiyitibwa punctate yellow fluorescence ku luwuzi lw’obutoffaali bwa kookolo ku ddakiika 30, ekiraga nti peptide etaliiko kamogo yaliwo ku luwuzi lw’obutoffaali bwa kookolo era yeeyongera n’okusengejja obutoffaali bwa kookolo. Okwawukana ku ekyo, obutoffaali bwa MCF-10-2A obutakyusiddwa mu kusooka bwalaga ekitangaala ky’olususu ekya kyenvu ekifaanagana, naye oluvannyuma ne kibula. Obutafaananako butoffaali bwa kookolo, obutoffaali buno obutakyusiddwa bwasigala nga buwangaala. Kino kiraga nti PNC-27 ereeta okukutuka kw’olususu mu butoffaali bwa kookolo ng’ekola nga peptide yonna, okusinga akatundu [2]..
Okusiba ku HDM-2 okukola ensengekera z’obutuli:
PNC-27 erimu HDM-2-binding structural domain ne cell-penetrating peptide (CPP) pressequence, ekireetera okutondebwa kw’ensengekera z’obutuli nga zisiba ku HDM-2 ku luwuzi lw’obutoffaali, ekivaako tumor cell lysis ne necrosis. Okubalirira kw’amasoboza ag’ensengekera (conformational energy calculations) kwalaga nti PNC-27 ekola ekizibu ne HDM-2 mu mugerageranyo gwa 1:1, ng’omutendera ogukulembera gusonga wala okuva ku kizibu. Okunoonyereza kwa Immunoscanning electron microscopy kwalaga nti waliwo obutundutundu bwa zaabu obuwera 6 nm ne 15 nm obuwandiikiddwa ku mugerageranyo nga 1:1 mu butuli ku ngulu w’obutoffaali bwa kookolo obujjanjabiddwa ne PNC-27, ekiraga nti ebizibu bino bikulu mu nsengeka y’obutuli. Okwawukana ku ekyo, tewali buziba bwakolebwa mu control, untransformed fibroblasts ezajjanjabwa ne PNC-27 [3]..
Kookolo w’olubuto:
Peptide empya elwanyisa kookolo eya PNC-27 ereeta okufa kw’obutoffaali bwa kookolo w’olubuto lw’omuntu MiaPaCa-2 mu ngeri eyeesigama ku ddoozi, naye si butwa eri obutoffaali obwa bulijjo. Okunoonyereza kulaga nti PNC-27, bw’egattibwa ne Gemzar, eddagala eriweweeza ku bulwadde bwa kookolo w’olubuto, lirina akakwate akasingako ku butoffaali bwa MiaPaCa-2 okusinga Gemzar oba PNC-27 yokka. Kino kiyinza okuba nga kiva ku kuba nti obutuli obuleetebwa PNC-27 byongera okuyita kw’obutoffaali bwa kookolo okutuuka ku Gemzar, ne kisobozesa Gemzar nnyingi okutunuulira obutoffaali bwa kookolo [1]..
Kookolo w’enkwaso:
Mu layini z’obutoffaali bwa kookolo w’enkwaso SKOV-3 ne OVCAR-3, puloteyina ya HDM-2 yeeyoleka nnyo ku luwuzi lw’obutoffaali.PNC-27 ekwatagana ne HDM-2 ku luwuzi lw’obutoffaali, ekivaako okufa kw’obutoffaali okw’amangu. Okwawukana ku ekyo, mu layini y’obutoffaali obufuga obutakyusiddwa HUVEC, co-localization ne cytotoxicity ya PNC-27 teyalabibwa olw’okwolesebwa kwa HDM-2 okw’olususu okutono [1]..
Kookolo w’olubuto olunene:
PNC-27 elonda okutta obutoffaali obusibuka mu kookolo w’olubuto olunene. Mu layini z’obutoffaali bwa kookolo w’olubuto mukaaga, PNC-27 yakwatagana ne membrane HDM-2 era n’ereeta okufa kw’obutoffaali (tumor cell necrosis, high LDH release, membrane associated protein V and cysteine 3 negativity) mu butoffaali bwa kookolo bwokka. Mu vivo, PNC-27 ereeta necrosis of tumor nodules naye tekosa bitundu bya bulijjo [1]..
3. Okulongoosa mu ngeri y’okulwanyisa ebizimba
Okusiba ku Doxil:
Kizuuliddwa nti PNC27 peptide, nga targeting ligand, esobola okutumbula ennyo anti-tumor efficacy ya Doxil mu butoffaali bwa kookolo obulina HDM2. Ebipimo eby’enjawulo ebya PNC27 peptide byayingizibwa mu Doxil, era flow cytometry n’okwekenneenya confocal byakolebwa ku butoffaali bwa kookolo w’olubuto olunene (HDM2-positive) ne B16F0 melanoma (HDM2-negative). Ebivuddemu biraga nti PNC27-Doxil yalaga okutwalibwa okw’amaanyi mu butoffaali n’obutwa mu butoffaali bwa C26, so nga ebivuddemu bino tebyalabibwa mu butoffaali bwa B16F0. Okugeza, PNC27-Doxil (100 PNC27 peptide) yayongera nnyo ku bujjanjabi bwa Doxil awatali kukosa kusaasaana kwayo mu biramu mu bizimba bya C26 [4]..
4. Okujjanjaba kookolo w’enkwaso mu ngeri ey’okugatta n’eddagala lya paclitaxel
Paclitaxel ekozesebwa nnyo mu kujjanjaba obulwadde obubi obw’abakyala, naye etunuulira obutoffaali bw’ekizimba mu mutendera gwa M ogw’enzirukanya y’obutoffaali, ng’obutoffaali mu mitendera emirala buwangaala era nga buyinza okuvaako ekizimba okuddamu.PNC-27 ye peptide ekoleddwa okuva mu amino acids mu P53-MDM-2 binding domain etta layini z’obutoffaali bwa kookolo obw’enjawulo mu ngeri eyeesigama ku ddoozi. Obutoffaali bwa kookolo w’enkwaso ID8 obwalagibwa paclitaxel bwazuulibwa nga bulaga okweyongera kw’okwolesebwa kwa MDM-2 n’okweyongera okuwuliziganya eri PNC-27.Ekikolwa ky’obutwa mu bitundu by’omubiri ekya PNC-27 kyesigamye ku kusiba kwayo ku MDM-2, era okuziyiza MDM-2 kiziyiza ekikolwa eky’okutta ekya PNC-27. Ebifaananyi bya heterodrug eby’okugatta ddoozi byali bya kukwatagana, ekiraga nti waliwo ekikolwa eky’okukwatagana wakati wa PNC-27 ne paclitaxel [5] . Mu nkola ya kookolo w’enkwaso (ID8) mu lubuto, okugattako PNC-27 ku kugaba paclitaxel buli wiiki kyakendeeza nnyo okukula kw’ekizimba. Ebiwandiiko bino biraga enkolagana wakati wa PNC-27 ne paclitaxel, nga PNC-27 etunuulira paclitaxel ewangaala obutoffaali n’okulongoosa ebikolwa byayo eby’okulwanyisa ebizimba.
5. Okujjanjaba kookolo w’enkwaso (primary epithelial ovarian cancer).
Okunoonyereza kutaddewo obuwangwa obusookerwako obw’obutoffaali bwa kookolo w’enkwaso (epithelial ovarian cancer cells) obupya obwawulwa okuva mu balwadde abaakazuulibwa nga balina kookolo w’enkwaso (varian cystadenocarcinoma). Omu ku bo yali ya mucinous cystadenocarcinoma ate omulala yali wa high-grade papillary plasmacytoid carcinoma. Obulung’amu bw’obujjanjabi bwa PNC-27 bwakeberebwa mu bungi nga bakozesa omutindo gw’ekitangaala microscopic observation ne MTT cell proliferation assay wamu n’okupima lactate dehydrogenase (LDH). Ebivuddemu biraga nti PNC-27 yaziyiza okukula kw’obutoffaali bwa kookolo obusookerwako obw’omuntu obwali bwaakawulwa okuva mu kookolo w’enkwaso abiri mu ngeri eyeesigama ku ddoozi era nga ya butwa bwa cytotoxic gye bali. Control peptide PNC-29 teyaliiko kye yakola ku butoffaali bwa kookolo obusookerwako. Okugatta ku ekyo, PNC-27 era ya butwa mu butoffaali eri layini z’obutoffaali bwa kookolo w’enkwaso z’omuntu ezimaze ebbanga nga zimanyiddwa era ezigumira eddagala [6]..
Mu bufunze, PNC-27, nga peptide empya elwanyisa kookolo, eraga obusobozi obw’amaanyi okukozesebwa mu bujjanjabi bwa kookolo obw’enjawulo. Mu kitundu ky’obulwadde bwa leukemia, PNC-27 yalaga ebikolwa eby’okutta okulonda ku acute myeloid leukemia (okugeza, U937, OCI-AML3, ne HL60 cell lines) n’obutoffaali bwa K562 obubulamu p53, era yatuuka ku bikolwa ebikola obulungi ennyo eby’okulwanyisa ebizimba nga ereeta obutoffaali okufa n’okufulumya lactate dehydrogenase. Mu kujjanjaba kookolo w’enkwaso, okugatta PNC-27 ne paclitaxel kwalaga ekikolwa eky’okukwatagana, ekiyinza okuziyiza ennyo okukula kw’ebizimba era nga kikyalina omulimu ogw’okutta obutoffaali obugumira eddagala. Okugatta ku ekyo, okugatta PNC-27 n’eddagala lya liposomal (okugeza Doxil) nga ligand egenderera kiyinza okutumbula enkola ey’enjawulo n’obulungi bw’eddagala eriziyiza ebizimba eri obutoffaali bw’ekizimba obulina HDM2. Enkola yaayo ey’okuziyiza ku butoffaali obusookerwako obwa kookolo w’enkwaso (primary epithelial ovarian cancer progenitor cells) kyongera okukakasa omugaso gw’okuvvuunula mu bujjanjabi. Mu kiseera kino, PNC-27 eyingidde mu kugezesebwa okw’omutendera I, era okunoonyereza okulala kwetaagibwa okukakasa obukuumi bwayo n’obulungi bwayo obw’ekiseera ekiwanvu, okusobola okugitumbula ng’eky’okugonjoola ekiyiiya mu kujjanjaba kookolo.
Ebikwata ku Muwandiisi
Ebintu ebyogeddwako waggulu byonna binoonyerezeddwako, birongooseddwa era ne bikung’aanyizibwa kkampuni ya Cocer Peptides.
Omuwandiisi wa Scientific Journal
Sarafraz-Yazdi E munoonyereza akwatagana n’ebibiina ebiwerako, omuli NomoCan, New York Blood Center, SUNY Downstate Health Sciences University, ne New York Harbor VA Med Ctr. Enkolagana zino ziraga okwenyigira kwe mu kunoonyereza n’embeera z’obusawo ez’enjawulo.
Okunoonyereza kwe kukwata ku masomo ag’enjawulo nga Oncology, Medical Laboratory Technology, Pharmacology & Pharmacy, Biochemistry & Molecular Biology, ne Research & Experimental Medicine. Omulimu gwe mu bintu bino gulaga obukugu bwe obugazi n’okwewaayo kwe okutumbula okumanya mu bintu bino ebikulu ebya ssaayansi w’obusawo n’okunoonyereza. ewandiikiddwa mu kiwandiiko ekijuliziddwa mu kiwandiiko ekijuliziddwa [3].
