Title AKTIVACIJSKI STATUS NK STANICA I MAKROFAGA U
SUROGATIMA MOLEKULARNIH PODTIPOVA KARCINOMA
DOJKE Title (english) ACTIVATION STATUS OF NK CELLS AND MACROPHAGES
IN SURROGATE MOLECULAR SUBTYPES OF BREAST
CANCER Author Alma Starčević Mentor Damir Grebić (mentor)Mentor Tamara Gulić (komentor)Committee member Ilija Brizić (predsjednik povjerenstva)Committee member Natalija Dedić Plavetić (član povjerenstva)Committee member Manuela Avirović (član povjerenstva)Granter University of Rijeka Defense date and country 2023, Croatia Scientific / art field, BIOMEDICINE AND HEALTHCARE Scientific / art field, BIOMEDICINE AND HEALTHCARE Scientific / art field, BIOMEDICINE AND HEALTHCARE Universal decimal classificationUDC ) 61 - Medical sciences ThesaurusMESH - Medical Subject Headings )
Breast Neoplasms
Killer Cells, Natural
Macrophages
Abstract Cilj istraživanja: ispitati aktivacijski status NK stanica i orijentaciju makrofaga na sistemskoj i lokalnoj razini u surogatima molekularnih podtipova karcinoma dojke (luminalni A, luminalni B i trostruko negativan karcinom) u odnosu na kontrolnu skupinu. Materijali i metode: Uzorke tkiva tumora dojke dobivali smo na Klinici za radiologiju KBC-a Rijeka, a potom ih slali na Zavod za opću patologiju i patološku anatomiju, gdje smo postavili dijagnozu i odredili surogat molekularnih podtipova karcinoma dojke. Od imunoloških metoda koristili smo imunohistokemijsko obilježavanje antigena u tkivu dojke uklopljenim u parafin (CD56, CD68, CD163, CD206, IL-15, APAF-1) uz semikvantitativnu analizu obilježenih rezova. Metodom imunoflorescencije periferne mononuklearne stanice bolesnica oboljelih od karcinoma dojke i kontrolne skupine obilježavali smo stanične biljege CD3/CD56/ i NKp46, NKG2A, NKG2C, CD94, CD56/CD69/CD45 i CD14/ i CD80, CD86, CD83, CD206 i CD163 uz očitavanje protočnim citometrom. Metodom lančane reakcije polimeraze u stvarnom vremenu (RT-qPCR) određivali smo izražaj gena za perforin, TRAIL i Fas-L. Rezultati: Udio CD56⁺ stanica, CD68+ stanica, CD163+ stanica i broj IL-15⁺ stanica povećan je u trostruko negativnom karcinomu u usporedbi s kontrolnom skupinom, luminalnim A i luminalnim B podtipom. Učestalost biljega NKp46 i NKG2C na NK stanicama u suspenziji mononuklearnih stanica periferne krvi statistički je značajno veća u kontrolnoj skupini u odnosu na sva tri podtipa karcinoma dojke. Izražaj NKG2A statistički je značajno veći u trostruko negativnom karcinomu u odnosu na kontrolnu skupinu, luminalni A i B podtip. Glasnička RNK za perforin petorostruko je veća u luminalnom B podtipu, slijedi isti porast genskog izražaja za ispitane citolitičke medijatore TRAIL i Fas-L. Učestalost CD80+ stanica, CD83+ stanica i CD86+ stanica značajno je viša u kontrolnoj skupini u odnosu na sva tri podtipa karcinoma dojke. Zaključak: Povećana učestalost tumoru pridruženih makrofaga (engl. Tumor-associated macrophages, TAMs) uglavnom alternativno aktiviranih, promijenjen fenotip i smanjen aktivacijski status NK stanica odlika je ponašanja malignijeg karcinoma dojke. Analiza genskog izražaja citolitičkih medijatora (perforina, TRAIL-a i Fas-L) nishodno je regulirana u malignijem obliku karcinoma dojke.
Abstract (english) Aim: To determinate the activation status of natural killer cells and the orientation of macrophages at the systemic and local level in surrogates of molecular subtypes of breast cancer (luminal A, luminal B and triple negative cancer) in comparison to the control group. Materials and methods: Breast tissue samples were obtained from Department of Radiology, Clinical Hospital Center Rijeka, and prepare to be analyzed for cancer breast classification in the Department of General Pathology and Patologic Anatomy. Immunohistochemistry was used to investigate following antigens in breast tissue sections embedded in paraffin CD56, CD68, CD163, CD206, IL-15, APAF-1 with semiquantitative analysis. Peripheral blood mononuclear cells were labelled with cell markers CD3/CD56/NKp46 or NKG2A or NKG2C or CD94 or CD107a; CD56/CD69/CD45 and CD14/ and CD80, CD86, CD83, CD206 and CD163 in breast cancer patients and the control group and analyed by flow cytometer. Using the real-time polymerase chain reaction method (RT-qPCR), we determined the genes expression for perforin, TRAIL and Fas-L. Results: The proportion of CD56+ cells, CD68+ cells, CD163+ cells and the number of IL-15⁺ cells were increased in triple-negative carcinoma compared to the control group, luminal A and luminal B subtype. The frequency of NKp46 and NKG2C markers on NK cells in the suspension of peripheral mononuclear cells was statistically significantly higher in the control group compared to the groups based on molecular division. The expression of NKG2A is statistically significantly higher in triple-negative carcinoma compared to the control group, luminal A and B subtype. The messenger RNA for perforin was five times higher in the luminal B subtype, and the same trend of increasing gene expression was observed for TRAIL and Fas-L. The frequency of CD80+, CD83+ and CD86+ cells was significantly higher in the control group than in luminal A subtype, luminal B subtype and triple negative breast cancer. Conclusion: An increased frequency of TAMs, mainly alternatively activated, a changed phenotype and reduced activation status of NK cells is a characteristic of the behavior of more malignant breast cancer. Analysis of the gene expression of cytolytic mediators (perforin, TRAIL and Fas-L) is down-regulated in the more malignant form of breast cancer.
Keywords
karcinom dojke
makrofagi
NK stanice
tumorski mikrookoliš
Keywords (english)
breast cancer
macrophages
natural killer cells
tumor microenvironment
Language croatian URN:NBN urn:nbn:hr:184:807852 Promotion 2023 Project Number: uniri-pr-biomed-19-17 Study programme Title: Biomedicine Postgraduate (doctoral) study programme Type of resource Text File origin Born digital Access conditions Access restricted to students and staff of home institution Terms of use Created on 2023-09-14 11:21:18
