Research On Pathology And Molecular Pathology Of Inverting Growth And Early Infiltration In Bladder Urothelial Carcinoma

Objective:1.To study the relationship between the positive expression of PASM-Masson and type IV collagen of inverted and invasive basement membrane of bladder urothelial carcinoma and the invasion.2.Use immunohistochemistry to differentiate inverted urothelial papilloma of bladder from inverted urothelial carcinoma of bladder.3.To study the application of microcarcinoma embolus cocktail antibody double-staining method in the diagnosis of vascular metastasis of bladder urothelial carcinoma.4.To study the molecular pathological expression and significance of bladder urothelial carcinoma.Use PASM-Masson,type IV collagen and cocktail staining of microcarcinoma embolus to assist morphological diagnosis of inverted and invasive urothelial carcinoma,establish objective diagnostic criteria to help pathologists differentiate inverted urothelial carcinoma from invasive urothelial carcinoma and whether there is vascular metastasis or not.Method:326 cases of bladder inverted urothelial papilloma,bladder inverted urothelial carcinoma and bladder invasive urothelial carcinoma from January 2010 to December2018 in Fuzhou General Hospital of Nanjing Military Region were collected.Among them,73 cases were bladder inverted urothelial papilloma,77 cases were bladder inverted urothelial carcinoma and 176 cases were bladder invasive urothelial carcinoma.The selected cases were stained with HE,PASM-Masson,IV collagen,and Ki-67,P53,microcarcinoma embolus cocktail antibody staining and molecularpathological examination.The classification and diagnosis criteria of bladder urothelial carcinoma published in the 2016 edition of <Tumor Pathology and Genetics of Urinary System and Male Reproductive Organs> by WHO were used.Results:1.Compared with the inverted non-invasive urothelial carcinoma group,there was significant increase in the loss rate of PASM-Masson and IV collagen staining of the basement membrane of the invasive urothelial carcinoma group,and the difference was significant between the two groups(P<0.001).The integrity of PASM-Masson and collagen IV staining of the basement membrane of inverted urothelial papilloma group was higher than that of invasive urothelial carcinoma group,and the difference was significant between the two groups(P<0.001).There was no significant difference in PASM-Masson and IV collagen staining between inverted papilloma group and inverted non-invasive urothelial carcinoma group(P >0.05).2.Ki-67 staining showed that among inverted urothelial papilloma group only 11 cases were positive(15%)and the other 62 cases were negative(85%).The expression of Ki-67 in inverted non-invasive urothelial carcinoma group was relatively lower,among which 74 cases(96%)were positive and 3 cases(4%)were negative.And the expression of Ki-67 in invasive urothelial carcinoma was very significant,among which 75 cases were positive(99%)and 1 case was negative(1%).The expression rate of Ki-67 in papilloma,inverted carcinoma and invasive carcinoma increased gradually and statistical analysis showed that the difference was significant(P < 0.05).And the difference of pairwise comparison of the three groups was statistically significant.P53 staining result showed that 70 cases(96%)were positive and 3 cases(4%)were negative in the inverted urothelial papilloma group.73cases(95%)were positive and 4 cases(5%)were negative in the inverted non-invasive urothelial carcinoma group.175 cases(99%)were positive and 1 case(1%)was negative in the invasive urothelial carcinoma group.Statistical analysisshowed that there were significant differences in the expression of P53 in papilloma,inverted cancer and invasive cancer(P < 0.05);and the difference of pairwise comparison of the three groups was statistically significant.3.The detection rates of vascular vision of double immunohistochemical staining and HE staining of microcarcinoma embolus cocktail antibody(D2-40,CD34-CK)were 38%(40/106)and 20%(21/106)respectively.Statistical analysis showed that the positive distribution of cocktail antibody D2-40/CD34-CK double immunohistochemical staining was significantly different from that of HE staining in bladder urothelial carcinoma(P < 0.05),suggesting that cocktail staining could improve the detection rate of vascular cancer thrombus.And in the comparison of cocktail antibody D2-40/CD34-CK double immunohistochemical staining and HE staining,the Kappa value was 0.491,P < 0.05,which was between 0.4 and 0.75,showing good consistency.4.Using Fluorescence in situ hybridization(FISH)to detect chromosome 3,7,17 and P16 gene,the positive expression rates were 5%(1/20)in bladder inverted urothelial papilloma group,20%(11/54)in bladder inverted urothelial carcinoma group and 78%(86/111)in bladder invasive urothelial carcinoma group.In the comparison of four probes in bladder inverted urothelial papilloma group and bladder inverted urothelial carcinoma group,the difference was statistically significant,P <0.05.In the comparison of four probes in bladder inverted urothelial papilloma group and bladder invasive urothelial carcinoma group,the difference was statistically significant,P < 0.05.In the comparison of four probes in inverted urothelial carcinoma group and invasive urothelial carcinoma group,the difference was no statistically significant(P > 0.05).Conclusion:1.PASM-Masson staining and IV collagen staining can be used to differentiate inverted papilloma with inverted growth type from invasive carcinoma,but can't be used to differentiate inverted papilloma and inverted non-invasive urothelialcarcinoma(in daily work,it mostly depends on histomorphology to differentiate them),which can help pathologists to judge whether there is early invasion on the inverted urothelial carcinoma.2.The expression of Ki-67 and P53 in urothelial carcinoma is higher than that in papillary carcinoma,and the expression in invasive carcinoma is higher than that in non-invasive carcinoma,which can be used to assist differential diagnosis with HE morphology.3.Cocktail double immunohistochemistry is suitable for the diagnosis of vascular cancer thrombus.It can also confirm the vascular invasion seen in routine HE staining section,eliminate the possible false positive,and find some small vascular cancer thrombus which is difficult to observe under HE section.4.Fluorescence in situ hybridization(FISH)is useful in differentiating benign and malignant urothelial tumors.5.The combined detection of immunohistochemistry,special staining and molecular pathology in bladder urothelial lesions is helpful to the pathological diagnosis and differential diagnosis of different bladder urothelial lesions.