| Abstract | CILJ ISTRAŽIVANJA bio je ispitati ispoljenost receptora za hijaluronsku kiselinu, CD 44 i RHAMM-a, u tkivu raka endometrija, te usporediti odnos njihove ispoljenostis kliničko-patološkim parametrima kao što su dob, dubina invazije miometrija, prisutnost limfokapilarne invazije, prisutnost metastaza u regionalnim limfnim čvorovima, histološki stupanj tumora i FIGO stadij bolesti. Nadalje, cilj nam je bio ispitati korelaciju ispoljenosti CD 44 i RHAMM-a s ispoljenošću estrogenskih i progesteronskih receptora. Na kraju, da bi utvrdili prognostičko značenje ispoljenosti CD 44 i RHAMM-a, cilj nam je bio ispitati povezanost njihove ispoljenostis preživljenjem bolesnica.MATERIJALI I METODE: za ovo istraživanje upotrijebljeni su parafinski blokovi tkivaraka endometrija 104 bolesnice liječenih na Klinici za ginekologiju i porodništvo Kliničkog bolničkog centra u Rijeci u razdoblju od 2000. do 2006. godine. Retroskpektivno je odabran materijal samo onih bolesnica u kojih je učinjen potpuni staging bolesti, odnosno u kojih je učinjena histerektomijas obostranom adneksektomijom i zdjeličnom limfadenektomijom. Za potrebe ovog istraživanja uzet je materijal samo pacijentica kod kojih je postavljena završna dijagnoza raka endometrija tipa I (endometrioidni). Ispoljenost RHAMM i CD 44 analizirana je imunohistokemijskom metodom na materijalu koji je fiksiran u 10% formalinu i uklopljen u parafin. Za analizu ispoljenosti RHAMM upotrijebljeno je monoklonalno RHAMM protutijelo, a za analizu ispoljenosti CD 44 upotrijebljeno je monoklonalno CD 44 protutijelo. Za analizu ispoljenosti estrogenskih receptora upotrijebljeno je mišje monoklonalno estrogen α protutujelo, a za ispoljenost progesteronskih receptora upotrijebljeno je mišje progesteron protutijelo. V Ispoljenost RHAMM i CD 44 procijenjena je semikvantitativno, a ispoljenost estrogenskih i progesteronskih receptora procijenjena je također semikvantitativno Allred score metodom. REZULTATI: prosječna životna dob bolesnica našeg uzorka bila je 60 godina. Kod 63,5% bolesnica dubina invazije miometrija je bila manja od polovice debljine stijenke maternice, a kod 36,5% invazija je bila dublja. Limfokapilarna invazija je nađena u 44,2% bolesnica, metastaze u zdjeličnim limfnim čvorovima u 5,8% bolesnica, a u paraortalnim limfnim čvorovima u 1,9% bolesnica. Najveći broj bolesnica je bio u FIGO IA stadiju bolesti, točnije 59,6% bolesnica. Dobro diferencirane tumore imalo je 51,9% bolesnica, 36,5% imalo je umjereno diferencirane, a 11,5% imalo je slabo diferencirane tumore. CD 44 je pokazao visokistupanj ispoljenosti u tkivu raka endometrija, naime samo u 7,7% slučajeva njegova ispoljenostje bila negativna. RHAMM je bio negativan u 42,3% slučajeva. Kod 23 bolesnice (22,1%) nađena je istovremena jaka ispoljenost CD 44 i RHAMM. Estrogenski receptori su bili pozitivni u velikom broju uzoraka, točnije u 86,5%, a progesteronski su bili pozitivni čak u 92,3% uzoraka. Nismo našli statistički značajnu korelaciju ispoljenosti CD 44 i RHAMM s ispoljenošćuestrogenskih i progesteronskih receptora. Usporedbom ispoljenostiCD 44 i RHAMM s kliničko-patološkim parametrima nađena jestatistički značajna povezanost njihove ispoljenosti s dubinom invazije miometrija, prisutnošću limfokapilarne invazije, FIGO stadijem bolesti, a u slučaju RHAMM-a i s histološkim stupnjem tumora. Primjetili smo da je ispoljenost CD 44 lokalizirana na staničnoj membrani u svim slučajevima, ali u određenom broju slučajeva postoji i citoplazmatska ispoljenost CD 44. Usporedbom te skupine bolesnica s ostalima, otkrili smo da među njima postoji statistički značajno veći udio slučajeva s dubljom invazijom miometrija (p=0,006) i limfokapilarnom invazijom (p=0,020). Izdvojili smo podskupinu od 23 bolesnice (22,1%) u kojih je nađena istovremena jaka ispoljenost RHAMM i CD 44, te smo ih usporedili s VI ostalim bolesnicama obziromna kliničko-patološke karakteristike. Našli smo da je u toj podskupini znatno veći udio bolesnica s dubljom invazijom miometrija (p=0,024), limfokapilarnom invazijom (p=0,022) i citopazmatskom ispoljenošću CD 44 (p=0,004). Na kraju smo analizirali preživljenje bolesnica univarijatnom analizom i ustanovili da u našem uzorku bolesnica ispoljenost CD 44 značajno utječe na petogodišnje preživljenje. Međutim, multivarijatnom cox-ovom regresijskom metodom niti CD 44 niti RHAMM nisu se pokazali kao neovisni prognostički čimbenici. ZAKLJUČCI: naši rezultati su pokazali da CD 44 i RHAMM imaju značajnu ulogu u invaziji tumorskih stanica u okolno tkivo te smo dokazali povezanost ispoljenosti ovih molekula s različitim kliničko-patološkim parametrima, a za sada samo pretpostavljamo njihovu vrijednost kao prognostičke čimbenike, za čiju potvrdu će biti potrebna buduća istraživanja. |
| Abstract (english) | OBJECTIVES: the aim of this study was to investigate the expression of the receptors for hyaluronan acid, CD 44 and RHAMM, in tissue samples of endometrial cancer as well as the relation of thier expression with some clinicopathological parameters, such as patient's age, the depth of myometrial invasion, lymph space involvement, pelvic and paraortic lymph node metastasis, histological tumour grade and FIGO stage of disease. Furthermore, the aim was to investigate the correlation of CD 44 and RHAMM expression with the expression of estrogen and progesteron receptors. Finally, in order to evaluate the value of CD 44 and RHAMM as prognostic factors, we investigated the relation of their expression with patients survival. MATERIALS AND METHODS: for this study we used tissue samples of endometrial cancer from 104 patients who were admitted to the Clinic of gynaecology and obstetrics, at Clinical Hospital of the University of Rijeka during the period between the year 2000 and 2006. All patients underwent total hysterectomy with bilateral adnexectomy and pelvic lymhadenectomy (with or without paraaortal lymphadenectomy), in other words, we retrospectively selected only patients with a complete staging. Only patients with the diagnosis of endometrioid adenocarcinoma (type I) were selected. The expression of CD 44 and RHAMM was analyzed with immunohistochemistry on material fixated in 10% formalin and embedded in VIIIparafin. For the analysis of the CD 44 expression a monoclonal CD 44 antibody was used, while for the analyisis of RHAMM expression we used a monoclosnal RHAMM antibody. For the analysis of the estrogen receptors expression we used a mice monoclonal estrogen α antibody while for the progesteron receptors expression analysis we used a mice monoclonal progesteron antibody. The expression of CD 44 and RHAMM was assessed semiquantitatively. The expression of estrogen and progesteron receptors was assessed also semiquantitatively, using the Allred score method.RESULTS: the average patients age was 60 years. In 63.5% of patients the depth of myometrial invasion was less then half the thickness of uterine wall, while in 36.5% of patients the invasion was deeper. Lymph space involvement was present in 44.2% of patients, we found pelvic lymph node metastasis in 5.8% of patients and paraortal lymph node metastasis in 1.9% of patients. The FIGO stage of disease for most of the patients was IA, precisely 59.6%. Well differentiated tumours were found in 51.9% of patients, moderatly differentiated tumours in 36.5% of patients and poorely differentiaed tumours in 11.5% of patients. CD 44 showed a high degree of expression in tissue samples of endometrial cancer, while in only 7.7% of cases its expression was negative. RHAMM expression was negative in 42.3% of cases. Interestingly, 23 patients (22.1%) showed a contemporaneously CD 44 and RHAMM strong expression. Estrogen receptors were positive in 86.5% of cases, while progesteron receptors were positive in 92.3% of cases. We didn't find a statistically significant correlation between CD 44 and RHAMM expression with estrogen and progesteron receptors. After we compared the CD 44 and RHAMM expression with clinicopathological findings we found a statistically significant correlation with the depth of myometrial invasion, lymph space involvement, FIGO stage of disease and in case of RHAMM expression also a significant correlation with the histological tumour grade. We IX noticed the CD 44 expression was localized in the tumour cell membrane in all cases, but in some cases the expression was positive in the cytoplasm of tumour cells as well. We compared this group of patients with the others and found a statistically significant increase of cases with deeper myometrial invasion (P =0.006) and lymph space involvement (P=0.020) within this group of patients. In addition to it, we sorted out the group of 23 (22.1%) patients with a contemporaneously strong CD 44 and RHAMM expression and compared them with the other patients regarding clinicopathological findings. In this group of patients a greater number of cases with deeper myometrial invasion (P=0.024), lymph space involvement (P=0.022) and cytoplasmatic CD 44 expression (P=0.004) was found. Finally we analysed patients survivalusing an univariate analyisis and found that in our sample of patients CD 44 expression showes a statistically significant influence on patients 5-year survival. But using a multivariate cox regression analyisis neither CD 44 or RHAMM confirmed themselfs as independent prognostic factors.CONCLUSIONS: our results showed that CD 44 and RHAMM have an important role in tumour invasion. Further, we demonstrated a correlation of CD 44 and RHAMM expression with various clinicopathological findings. For now we can only assume thier value as prognostic factors and to asses their exact value as prognostic markers for patients with endometrial cancer further investigations should be done. |
