Kuyezetsa DNA methylation pamodzi ndi mafoni a m'manja kuti ayesedwe koyambirira kwa zotupa ndi kuyezetsa khansa ya m'magazi ndi kulondola kwa 90.0%!

Kuzindikira khansa msanga pogwiritsa ntchito njira yodziwira khansa ndi njira yatsopano yodziwira khansa yomwe yaperekedwa ndi US National Cancer Institute m'zaka zaposachedwa, ndi cholinga chozindikira khansa yoyambirira kapena ngakhale zilonda zomwe zisanachitike khansa. Yagwiritsidwa ntchito kwambiri ngati chizindikiro chatsopano chodziwira khansa yoyambirira, kuphatikizapo khansa ya m'mapapo, zotupa zam'mimba, ma gliomas ndi zotupa za amayi.

Kupezeka kwa mapulatifomu ozindikiritsa zizindikiro za methylation landscape (Methylscape) kungathandize kwambiri kuwunika khansa koyambirira, zomwe zimapangitsa odwala kukhala pamlingo woyambirira wochiritsika.

Posachedwapa, ofufuza apanga njira yosavuta komanso yolunjika yodziwira malo a methylation pogwiritsa ntchito tinthu tating'onoting'ono tagolide tokongoletsedwa ndi cysteamine (Cyst/AuNPs) tophatikizidwa ndi biosensor yochokera ku mafoni yomwe imalola kuyeza msanga mitundu yosiyanasiyana ya zotupa. Kuyezetsa msanga kwa khansa ya m'magazi kumatha kuchitika mkati mwa mphindi 15 DNA itachotsedwa m'magazi, ndi kulondola kwa 90.0%. Mutu wa nkhaniyi ndi Kuzindikira mwachangu DNA ya khansa m'magazi a anthu pogwiritsa ntchito cysteamine-capped AuNPs ndi foni yam'manja yothandizidwa ndi makina ophunzirira.

Chithunzi 1. Njira yosavuta komanso yofulumira yodziwira khansa kudzera mu zigawo za Cyst/AuNPs ingathe kuchitika m'njira ziwiri zosavuta.

Izi zikuwonetsedwa pa Chithunzi 1. Choyamba, yankho lamadzi linagwiritsidwa ntchito kusungunula zidutswa za DNA. Kenako ma Cyst/AuNPs anawonjezeredwa ku yankho losakanikirana. DNA yachibadwa ndi yoyipa ili ndi mawonekedwe osiyanasiyana a methylation, zomwe zimapangitsa zidutswa za DNA kukhala ndi mitundu yosiyanasiyana yodzipangira yokha. DNA yachibadwa imasonkhana momasuka ndipo pamapeto pake imasonkhanitsa ma Cyst/AuNPs, zomwe zimapangitsa kuti Cyst/AuNPs isinthe kukhala yofiira, kotero kuti kusintha kwa mtundu kuchokera ku wofiira kupita ku wofiirira kungathe kuwonedwa ndi maso amaliseche. Mosiyana ndi zimenezi, mawonekedwe apadera a methylation a DNA ya khansa amachititsa kuti pakhale magulu akuluakulu a zidutswa za DNA.

Zithunzi za mbale 96 za zitsime zinajambulidwa pogwiritsa ntchito kamera ya foni yam'manja. DNA ya khansa inayesedwa ndi foni yam'manja yokhala ndi makina ophunzirira poyerekeza ndi njira zogwiritsira ntchito spectroscopy.

Kuyezetsa khansa m'magazi enieni

Pofuna kukulitsa ntchito ya chipangizo chowunikira, ofufuzawo adagwiritsa ntchito sensa yomwe idasiyanitsa bwino pakati pa DNA yachibadwa ndi ya khansa m'magazi enieni. Mapangidwe a methylation m'malo a CpG amawongolera momwe majini amagwirira ntchito. Pafupifupi mitundu yonse ya khansa, kusintha kwa methylation ya DNA komanso momwe majini amagwirira ntchito zomwe zimayambitsa tumorigenesis kwawonedwa kuti kumasinthana.

Monga chitsanzo cha khansa zina zokhudzana ndi methylation ya DNA, ofufuzawa adagwiritsa ntchito zitsanzo za magazi kuchokera kwa odwala khansa ya m'magazi ndi owongolera athanzi kuti afufuze momwe methylation imagwirira ntchito posiyanitsa khansa ya m'magazi. Chizindikiro cha methylation ichi sichimangopambana njira zomwe zilipo zowunikira khansa ya m'magazi mwachangu, komanso chikuwonetsa kuthekera kofikira mpaka kuzindikira msanga mitundu yosiyanasiyana ya khansa pogwiritsa ntchito njira yosavuta komanso yosavuta iyi.

DNA yochokera m'magazi kuchokera kwa odwala 31 a khansa ya m'magazi ndi anthu 12 athanzi idawunikidwa. Monga momwe zasonyezedwera mu bokosi la Chithunzi 2a, kuyamwa kwa zitsanzo za khansa (ΔA650/525) kunali kotsika kuposa kwa DNA yochokera m'masampuli abwinobwino. Izi makamaka zinali chifukwa cha kuwonjezeka kwa hydrophobicity komwe kunapangitsa kuti DNA ya khansa isakanike kwambiri, zomwe zinaletsa kusonkhana kwa Cyst/AuNPs. Zotsatira zake, tinthu tating'onoting'ono timeneti tinafalikira kwathunthu m'magawo akunja a maselo a khansa, zomwe zinapangitsa kuti Cyst/AuNPs zisakanike mosiyana pa maselo abwinobwino ndi a khansa. Ma curve a ROC adapangidwa posintha malire kuchokera pamtengo wotsika wa ΔA650/525 mpaka mtengo wapamwamba kwambiri.

Chithunzi 2.(a) Kuchuluka kwa kuyamwa kwa ma cyst/AuNPs omwe akuwonetsa kukhalapo kwa DNA yabwinobwino (yabuluu) ndi khansa (yofiira) pansi pa mikhalidwe yabwino.

(DA650/525) ya ma box plots; (b) Kusanthula kwa ROC ndi kuwunika mayeso ozindikira matenda. (c) Matrix yosokoneza kuzindikira odwala abwinobwino ndi odwala khansa. (d) Kuzindikira, kutsimikiza, kuneneratu kwabwino (PPV), kuneneratu koipa (NPV) ndi kulondola kwa njira yomwe yapangidwa.

Monga momwe zasonyezedwera pa Chithunzi 2b, dera lomwe lili pansi pa ROC curve (AUC = 0.9274) lomwe linapezedwa pa sensa yopangidwayo linasonyeza kukhudzidwa kwakukulu ndi kulunjika. Monga momwe zikuonekera kuchokera mu bokosilo, mfundo yotsika kwambiri yomwe ikuyimira gulu la DNA labwinobwino silimasiyana bwino ndi mfundo yapamwamba kwambiri yomwe ikuyimira gulu la DNA la khansa; chifukwa chake, logistic regression idagwiritsidwa ntchito kusiyanitsa pakati pa magulu abwinobwino ndi a khansa. Popeza pali mitundu yosiyanasiyana yodziyimira payokha, imayesa kuthekera kwa chochitika kuchitika, monga khansa kapena gulu labwinobwino. Zosintha zomwe zimadalira zimakhala pakati pa 0 ndi 1. Chifukwa chake, zotsatira zake ndi kuthekera. Tinapeza kuthekera kwa kuzindikira khansa (P) kutengera ΔA650/525 motere.

kumene b=5.3533,w1=-6.965. Pakugawa zitsanzo, mwayi wochepera 0.5 umasonyeza chitsanzo chabwinobwino, pomwe mwayi wa 0.5 kapena kupitirira apo umasonyeza chitsanzo cha khansa. Chithunzi 2c chikuwonetsa matrix ya chisokonezo yopangidwa kuchokera ku kutsimikizira kwa leave-it-alone, komwe kunagwiritsidwa ntchito kutsimikizira kukhazikika kwa njira yogawa. Chithunzi 2d chikufotokoza mwachidule kuwunika kwa mayeso ozindikira njira, kuphatikiza kukhudzidwa, kutsimikizika, mtengo woneneratu wabwino (PPV) ndi mtengo woneneratu woipa (NPV).

Masensa a biosensor opangidwa ndi mafoni a m'manja

Kuti apititse patsogolo kuyesa zitsanzo popanda kugwiritsa ntchito ma spectrophotometer, ofufuzawo adagwiritsa ntchito luntha lochita kupanga (AI) kuti afotokoze mtundu wa yankho ndikusiyanitsa pakati pa anthu abwinobwino ndi omwe ali ndi khansa. Popeza izi zidachitika, masomphenya apakompyuta adagwiritsidwa ntchito kumasulira mtundu wa yankho la Cyst/AuNPs kukhala DNA yachibadwa (wofiirira) kapena DNA ya khansa (yofiira) pogwiritsa ntchito zithunzi za mbale 96-well zomwe zidatengedwa kudzera mu kamera ya foni yam'manja. Luntha lochita kupanga lingachepetse ndalama ndikuwonjezera mwayi wopeza mtundu wa mayankho a nanoparticle, komanso popanda kugwiritsa ntchito zida zilizonse zamafoni. Pomaliza, mitundu iwiri yophunzirira makina, kuphatikiza Random Forest (RF) ndi Support Vector Machine (SVM) idaphunzitsidwa kuti ipange mitunduyi. Mitundu yonse ya RF ndi SVM idagawa bwino zitsanzozo ngati zabwino ndi zoyipa ndi kulondola kwa 90.0%. Izi zikusonyeza kuti kugwiritsa ntchito luntha lochita kupanga poyesa kugwiritsa ntchito mafoni ndizotheka.

Chithunzi 3.(a) Gulu la yankho lolembedwa panthawi yokonzekera chitsanzo cha gawo lopezera chithunzi. (b) Chitsanzo cha chithunzi chomwe chinatengedwa panthawi yopezera chithunzi. (c) Kulimba kwa utoto wa yankho la cyst/AuNPs mu chitsime chilichonse cha mbale ya zitsime 96 yotengedwa pachithunzichi (b).

Pogwiritsa ntchito Cyst/AuNPs, ofufuza apanga bwino njira yosavuta yodziwira malo a methylation komanso sensa yokhoza kusiyanitsa DNA yabwinobwino ndi DNA ya khansa pogwiritsa ntchito zitsanzo zenizeni za magazi poyesa khansa ya m'magazi. Sensa yopangidwayo yawonetsa kuti DNA yotengedwa kuchokera ku zitsanzo zenizeni za magazi inali yokhoza kuzindikira mwachangu komanso mopanda mtengo kuchuluka kwa DNA ya khansa (3nM) mwa odwala khansa ya m'magazi mumphindi 15, ndipo inasonyeza kulondola kwa 95.3%. Kuti tiwonjezere kusavuta kuyesa zitsanzo mwa kuchotsa kufunikira kwa spectrophotometer, kuphunzira kwa makina kunagwiritsidwa ntchito kutanthauzira mtundu wa yankho ndikusiyanitsa pakati pa anthu abwinobwino ndi odwala khansa pogwiritsa ntchito chithunzi cha foni yam'manja, ndipo kulondola kunathekanso kupezeka pa 90.0%.

Chizindikiro: DOI: 10.1039/d2ra05725e