Angiogeneza cervikalnih intraepitelnih neoplazija (CIN) : kvantitativna morfometrijska analiza [ Angiogenesis of cervical intraepithelial neoplasia (CIN) : quantitative morphometric analysis ]

Mrčela, Milanka (2005) Angiogeneza cervikalnih intraepitelnih neoplazija (CIN) : kvantitativna morfometrijska analiza [ Angiogenesis of cervical intraepithelial neoplasia (CIN) : quantitative morphometric analysis ]. PhD thesis, Sveučilište u Zagrebu.

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Abstract

The aim of this research was to answer whether cervical intraepithelial neoplasias are able to provoke neovascularisation - angiogenesis. The quantity (the sum of micro-vessels along with the basement membrane of epithelium and total number of vessels of set microscopic view field) and the quality of vessels (diameter, circumference and area) were compared between 30 specimens of CIN 1, 30 specimens of CIN 2, 30 specimens of CIN 3 and two control groups each of them containing 30 specimens of cervical tissue without any pathohistological abnormality and differing from each other in the fact that specimens of control group K 1 were taken from patients who underwent hysterectomy for reasons unrelated to CIN and group K 2 whose specimens were taken from patients with CIN 3 lesions but taken from parts wich had no pathohistological abnormality. Samples were treated with histochemical methods PAS and Mallory and with immunohistochemical methods with anti-F-VIII and anti-CD 31 markers. Quantitative and qualitative parameters were processed using quantitative morphometric analysis in program "ISSA for windows", produced by VAMS, Zagreb. Classic histological criteria such as degree of inflammation, the CIN cell size, the number of CIN mitoses and the presence of koilocytes were all processed by semiquantitative approach and compared with the results of quantitative morphometrics. The results were statistically analysed by method testing analysis of variance (ANOVA) based on Fischer' s (F-test) test as the criterion of significance. No significant difference between control groups K 1 and K 2 (P>0,05) was found. Statistically significant difference was found in the sum of micro-vessels directly below or along the basement membrane between CIN 1, CIN 2, CIN 3 and the control group K 1 using immunohistochemical methods F-VIII, CD 31 and histochemical method Mallory (P<0,05) and between CIN 3 and the control group K 1 in all aplied methods (P<0,05). A reliable correlation between CIN grades and vessels quality was not found. The number of vessels close to the basement membrane of epithelium between CIN 1 and CIN 3. and CIN 2 and CIN 3 in both immunohistochemical methods was significantly different (P F-VIIi < 0,05 and P CD31 <0,05). The correlation between inflammation degree, CIN cell size, number of CIN mitoses and quality and quantity of blood vessels was not found. However, the presence of koilocytes affects the number of blood vessels along the basement membrane of dysplastic epithelium. Since there is a significant difference in the number of blood vessels along the basement membrane between normal and dysplastic epithelium and the number increases with the rise of CIN degree, it was concluded that CIN lesions are angiogenic.

Abstract in Croatian

Ovim istraživanjem nastojalo se odgovoriti na pitanje jesu li cervikalne neoplazije sposobne izazvati neovaskularizaciju - angiogenezu. Kvantiteta (zbroj krvnih žila uz bazalnu membranu epitela i ukupan broj krvnih žila zadanog vidnog polja) te kvaliteta krvnih žila (promjer, opseg i površina) uspoređivani su između 30 uzoraka CIN 1, 30 uzoraka CIN 2, 30 uzoraka CIN 3 te dviju kontrolnih skupina od kojih svaka sadrži po 30 uzoraka tkiva cerviksa bez patohistološke abnormalnosti, a medđusobno se razlikuje po tome što su uzorci skupine K 1 uzeti s cerviksa bolesnica histerektomiranih zbog razloga koji nemaju veze s CINom, a uzorci skupine K 2 od bolesnica s CIN 3, ali s mjesta bez patohistološke abnormalnosti. Uzorci su histokemijski obrađeni po PASu i Malloryu te imunihistokemijski metodama s biljezima anti-F-VIII i anti-CD31. Kvantitativni i kvalitativni parametri obrađeni su kvantitativnom morfometrijskom analizom u programu "ISSA for windows" firme VAMS, Zagreb. Klasični histološki pokazatelji kao stupanj upale, veličina CINe, broj mitoza CINe i prisustvo koilocita obrađeni su polukvantitativnim pristupom i uspoređeni s rezultatima kvantitativne morfometrije. Rezultati su statistički obrađeni testiranjem analize varijance metodom (ANOVA), a kriterij značajnosti bio je Fischerov F-test. Značajke kvantitativne i kvalitativne razlike između krvnih žila kontrolnih skupina K1 i K2 nisu nađene (P>0.05). Nađena je statistički značajna razlika u broju krvnih žila neposredno ispod ili uz bazalnu membranu epitela između CIN 1, CIN 2, CIN 3 i kontrolne skupine K 1 u imnunohistokemijskim metodama F-VIII i CD 31 i metodi Mallory (P<0,05) te između CIN 3 i kontrolne skupine K 2 u svim primjenjenim metodama (P< 0,05). Statistički značajna razlika u ukupnom broju krvnih žila nađena je samo između kontrolnih skupina K 1 i K 2 i CIN 3 u metodi F-VIII (P<0,05). Pouzdana korelacija između stupnja CINe i kvaliteta krvnih žila nije nađena. Broj krvnih žila neposredno uz bazalnu membranu epitela između CIN 1 i CIN 3, te između CIN 2 i CIN 3 u obje imunohistokemijske metode statistički se značajno razlikuje (P F-VIIi < 0,05 i P CD31 <0,05). Korelacija stupnja upale, veličine stanica CINe, broja mitoza CINe s kvalitetom i kvantitetom krvnih žila CINa nije nađena. Međutim, prisustvo koilocita utječe na broj krvnih žila uz bazalnu membranu displastičnog epitela. Budući postoji značajna razlika u broju krvnih žila uz bazalnu membranu između normalnog i displastičnog epitela, a taj broj raste s porastm stupnja CINe, došlo se do zaključka da su cervikalne intraepitelne neoplazije angiogene.

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