| Abstract | Cilj istraživanja. Analizirati povezanost izražaja katepsina K u malignom melanomu s morfološkim prognostičkim čimbenicima i ispitati njegov prognostički značaj.
Ustroj istraživanja. Prikupljen je bioptički materijal ispitanika s malignim melanomom kože (n=45) dijagnosticiranim u razdoblju od 1. siječnja 2005. godine do 1. prosinca 2011. godine. Uključujući kriteriji bili su potpuno odstranjena primarna lezija, potpuni klinički staging nakon dijagnoze malignog melanoma i redovito kliničko praćenje vremena do pojave metastaza. Nakon imunohistokemijskog bojenja analiziran je izražaj katepsina K i ispitana povezanost s predskazateljima nepovoljne prognoze malignog melanoma.
Mjesto istraživanja. Klinički zavod za patologiju, sudsku medicinu i citologiju Kliničkog bolničkog centra Split; Split; Hrvatska.
Tvoriva i postupci. Istraživanje je provedeno na bioptičkom materijalu malignog melanoma kože dijagnosticiranog u 45 ispitanika. Iz pismohrane su prikupljeni podatci o dobi i spolu ispitanika te pojavi metastaza u limfne čvorove i udaljene organe, a iz arhive uzeti originalni histološki preparati obojeni hemalaun-eozinom i parafinski blokovi. Na histološkim preparatima su analizirani: histološki tip tumora, dubina invazije tumora prema Clarku, debljina tumora prema Breslowu, površina tumora u mm², prisutnost ulceracije, pagetoizacije, patoloških mitotskih figura, upalne infiltracije, limfokapilarne invazije i mikrosatelitskih čvorova. U laboratoriju za imunohistokemiju izrezani su parafinski blokovi i obojeni na Anti-Cathepsin K (Abcam, Cambridge, Velika Britanija). Rezultati bojenja analizirani su svjetlosnim mikroskopom. U statističkoj obradi podataka korišteni su χ² test, Mann-Whitney test i Fisherov egzaktni test, a Kaplan-Meierovom krivuljom prikazano je prosječno vrijeme do pojave metastaza u limfne čvorove i udaljene organe.
Glavne mjere ishoda. Prosječno vrijeme bez pojave bolesti (engl. Disease-Free Survival – DFS), odnosno vrijeme do pojave metastaza u limfne čvorove i udaljene organe, promatrano od 1. siječnja 2005. godine do 1. prosinca 2011. godine i prikazano Kaplan-Meierovom krivuljom.
Rezultati. Nije utvrđena statistički značajna razlika izražaja katepsina K po spolu niti po životnoj dobi (P=0,736; P=0,118). U skupini ispitanika s pozitivnim izražajem katepsina K je 2,7 puta više nodularnog melanoma u odnosu na lentigo maligna i površinsko šireći melanom (P=0,014). U skupini ispitanika s pozitivnim izražajem katepsina K je gotovo 7 puta više ispitanika sa stupnjem invazije IV i V prema Clarku u odnosu na ispitanike sa stupnjem invazije I, II i III prema Clarku (P<0,001), a gotovo 5 puta više ispitanika s debljinom tumora stadija III, IV i V prema Breslowu u odnosu na ispitanike s debljinom tumora stadija I i II prema Breslowu (P<0,001). Statistički značajnu povezanost s pozitivnim izražajem katepsina K pokazala je prisutnost ulceracije (P=0,008), a statistički značajnu povezanost s negativnim izražajem katepsina K pokazala je prisutnost pagetoizacije (P=0,003). Veći broj mitoza na 10 vidnih polja velikog povećanja i veća površina tumora u mm2 su statistički značajno povezane s pozitivnim izražajem katepsina K (P=0,001; P<0,001). Prosječno vrijeme do pojave metastaza u limfne čvorove i udaljene organe izraženo Kaplan-Meierovom krivuljom statistički je značajno povezano s pozitivnim izražajem katepsina K (P<0,001).
Zaključci. Pozitivan izražaj katepsina K u stanicama malignog melanoma je značajno povezan s nepovoljnim morfološkim prognostičkim čimbenicima i kraćim vremenom do pojave metastaza u limfne čvorove i udaljene organe te bi njegov neovisni nepovoljni prognostički značaj trebalo ispitati na većim serijama. |
| Abstract (english) | Objective. To analyse the connection between the expression of cathepsin K in malignant melanoma and the morphological prognostic factors with its prognostic value.
Design. The research was conducted on biopsy material of 45 patients diagnosed with malignant melanoma, observed since 1st January 2005 until 1st December 2011. Including criteria were completely removed primary lesion, complete clinical staging after the diagnosis of malignant melanoma and regular clinical follow-up time to the occurrence of metastases. Expression of cathespin K was analyzed after immunostaging. Expression of cathepsin K was analyzed in corelation with morfological prognostic factors of malignant melanoma.
Settings. Institute for Clinical Pathology, Forensic Medicine and Cytology at the Clinical Hospital Centre Split; Split, Croatia.
Materials and Methods. The research was conducted on biopsy material of 45 patients diagnosed with malignant melanoma. The data about the age and sex of the patients along with the appearance of the metastasis was collected from medical documentation and the original histopathological preparations colored with hemalaun-eozin and the paraffin blocks were taken from the archive. The following was analyzed on the histopathological preparations: histopathological type of the tumor, depth of invasion (Clark's levels), Breslow's thickness and the surface of the tumor measured in mm², the presence of ulceration, Paget's cells resembling, inflammatory infiltrations, lymphovascular invasion and tumor's microsatellites. Immunostaining was performed on paraffin-embedded sections of 45 malignant melanomas. χ² test, Mann-Whitney test and Fisher's exact test were used in the statistical evaluation of the data. Disease Free Survival (DFS), time without the appearance of metastases in lymph nodes and distant organs, was shown with the Kaplan-Meier curve.
Main Outcome Measures. Disease Free Survival – DFS, time without the appearance of metastases in lymph nodes and distant organs, observed since 1st January 2005 until 1st December 2011, and shown with the Kaplan-Meier curve.
Results. A statistically significant difference in the expression of cathepsin K was not identified whether with sex or age (P=0,736; P=0,118). Within the group of patients with a positive expression of cathepsins K nodular melanoma was found 2,7 times more than lentigo maligna and superficial spreading melanoma (P=0,014). According to Clark's level, in the group of patients with a positive expression of cathepsins K, we have almost 7 times more patients with an invasion stage IV and V in reference to the examinees with an invasion stage I, II and III (P<0,001) and, according to Breslow, almost 5 times more patients with a tumor stage III, IV and V in reference to the patients with a tumor stage I and II (P<0,001). A statistically notable association with the positive expression of cathepsin K was shown while analyzing ulcerations (P=0,008), but statistically notable association with the negative expression of cathepsin K was shown while analyzing Paget's cells resembling (P=0,003). A larger number of mitosis on 10 fields of view of great enlargement and a larger surface of the tumor measured in mm2 are significantly associated with the positive expression of cathepsin K (P=0,001; P<0,001). The average time until the appearance of metastases in lymph nodes and distant organs expressed with the Kaplan-Meier curve is significantly associated with the positive expression of cathepsin K.
Conclusions. The positive expression of cathepsin K in malignant melanoma cells is significantly associated with the adverse morphological prognostic factors and a shorter Disease Free Survival (DFS). Its independent adverse prognostic value has to be examined on a greater scale of series. |
