ROLE OF TUMOR-ASOCIATED GLYCOPROTEIN-72 IN ENDOMETRIAL CARCINOMA

Cilj istraživanja: kvantificirati prisutnost CK (cytokeratin) 19 and TAG-72 (engl. tumor associated glycoprotein 72) u raku endometrija i usporediti rezultate s imunološkim svojstvima tumorskog mikrookoliša i kliničko-patološkim prognostičkim čimbenicima. Material i metode: U istraživanje je uključeno 107 uzoraka endometriodinog raka endometrija od kojih su izrađeni tkivni mikroareji (eng. Tissue microaray TMA). Metodom imunohistologije obilježavali smo CK 19, TAG-72, CD68, arginazu-1, iNOS (engl. inducible nitric oxide synthase) i Apaf-1 (engl. apoptotic protease activating factor 1) te kvantificirali broj stanica i jačinu obilježavanja. Fluorescentnim mikroskopom analizirali smo polarizaciju makrofaga [CD68/interleukin (IL)-15, CD68/CC ligand (CCL)2, CD68/arginaza-1, CD68/CCL22], prostorni međuodnos biljega [CK19/TAG-72, CD68/CD56, CK19/perforin, CD19/TRAIL (TNF-related apoptosis-inducing ligand)]. Rezultati:U endometrioidnom raku endometrija preko 90% stanica je izražavalo TAG-72 i CK19. Granularni izražaj TAG-72 pozitivno je korelirao s mucinoznom komponentom. Difuzno membranski i citoplaznatski izražaj TAG-72 u endometrioidnom raku endometrija s mucinoznom komponentom pozitivno je korelirao s brojem CD68+ stanica u centru tumora, dok je kontinuirano membranski izražaj TAG-72 u tumorskim žljezdanim stanicama pozitivno korelirao s veličinom tumora i histološkim gradusom tumora, a negativno s preživljenjem bolesnica. U centru tumora i na invazivnoj fronti endometrioidnog raka endometrija s mucinoznom komponentom CD68+ makrofazi izražavaju TAG-72 i posjeduju arginaza- 1+CCL22+CCL2- i IL-15 fenotip s obilježjima M2 aktivacije. Tkivno uvjetovane CD68+stanice okružuju rijetke NK stanice siromašne s citolotičkim medijatorom perforinom i apoptotičkom molekulom TRAIL, a broj nekrotičnih tumorskih CK19+ stanica s perforinom u citoplazmi i broj Apaf-1+ apoptotičnih stanica je bio vrlo mali. Endometrioidni rak endometrija bez mucinozne komponente je bio infiltran CD68+ makrofagima, koji nisu izražavali agrinazu-1, CCL22, ali su stvarali CCL2 i IL- 15 te imali veći udio CD56+ stanica i Apaf-1 pozitivnih apoptotičnih stanica. Zaključak: TAG-72 obilno izražen na stanicama endometrioidnog raka endometrija s mucinoznom komponentom podržava M2 program sazrijevanja makrofaga i osposobljava ih za podešavanje lokalne tolerogene imunološke reakcije djelovanjem na citotoksični potencijal stanica NK, što bi moglo podržavati nekontrolirani lokalni rast tumora i skratiti preživljenje bolesnica.Aim of the study: To quantify the presence of CK (cytokeratin) 19 and TAG-72 (tumor associated glycoprotein 72) in endometrial cancer and to compare the results with the immunological properties of the tumor microenvironment and clinical-pathological prognostic factors. Material and methods: Tissue microarrays (TMA) were built form 107 archival endometriod endometrial cancers. Immunohistology was used to label cytokeratin 19, TAG-72, CD68, arginase-1, iNOS (inducible nitric oxide synthase) and Apaf-1 (apoptotic protease activating factor 1) and to quantify cell number and labeling strength. Fluorescence microscopy was used to analyze the polarization of macrophages [CD68/interleukin (IL)-15, CD68/CC ligand (CCL) 2, CD68/arginase-1, CD68/CCL22], spatial interrelationship of markers [CK19/TAG-72, CD68/CD56, CK19/perforin, CD19/TRAIL (TNF-related apoptosis-inducing ligand)]. Results: Over 90% of cells expressed TAG-72 and CK19. Granular expression of TAG-72 was positively correlated with mucinous component. Diffuse membrane and cytoplasmic expression of TAG-72 in endometrioid endometrial cancer with mucinous component was positively correlated with the number of CD68+ cells in the tumor center; continous membrane expression of TAG-72 in tumor glandular cells was positively correlated with tumor size and histological grade and negatively with the overall survival of patients. In tumor center and invasive front of endometrioid endometrial cancer with mucinous component CD68+ macrophages express TAG-72 and possess arginase-1+CCL22+CCL2- and IL-15 phenotype with M2 activation characteristics. Tissue-mediated CD68+ cells surround NK cells poor in the cytolytic mediator perforin and apoptotic molecule TRAIL, and number of necrotic tumor CK19+ cells with perforin in the cytoplasm and the number of Apaf-1+ aptotic cells were very small. Endometrioid endometrial cancer without a mucinous component was infiltrated with CD68+ macrophages, which did not express agrinase-1, CCL22, but produced CCL2 and IL-15, and had a higher proportion of CD56+ cells and Apaf-1 positive apoptotic cells. Conclusion: TAG-72 is in abundance expressed in endometrioid endometrial cancer cells with a mucinous component, supports the M2 macrophage maturation program which could support uncontrolled local tumor growth and shorten patient survival