脉络膜黑色素瘤脱色素处理后FⅧ-RAg、VEGF的免疫组化染色

The immunohistochemical staining of FⅧ-RAg and VEGF after melanin-bleaching in choroidal melanoma

目的 :观察脉络膜黑色素瘤经两种脱色素法处理后第八因子相关抗原 (factorⅧ -relatedantigen ,FⅧ -RAg)、血管内皮生长因子 (vascularendothelialgrowthfactor ,VEGF)的表达情况 ,选择在脉络膜黑色素瘤血管发生学研究中进行免疫组化染色的适宜的脱色素方法。方法 :66例脉络膜黑色素瘤石蜡组织标本 ,改良高锰酸钾 /草酸溶液脱色素法 ,并与 10 %H2 O2 脱色素法相对照 ,应用Envision免疫组化法检测FⅧ -RAg、VEGF蛋白的表达。结果 :所有切片均呈不同程度的FⅧ -RAg、VEGF蛋白表达 ,且不受脱色素的影响。高锰酸钾 /草酸溶液脱色素法较 10 %H2 O2脱色素法具有操作简便、费时短 ,不易掉片 ,去除色素彻底等特点 ,而且保持了组织的抗原性。结论 :高锰酸钾 /草酸溶液脱色素法是脉络膜黑色素瘤免疫组化染色较为适宜的脱色素方法 ,对FⅧ -RAg、VEGF蛋白的表达无影响。

Objective:To investigate the effect of melanin-bleaching using permanganate/oxalate on subsequent immunohistochemical staining of FⅧ-Rag and VEGF in choroidal melanoma. Methods:66 cases of choroidal melanoma with different degree of pigmentation were selected.A modification of the potassium permanganate/oxalic acid melanin-bleaching technique was used,compared with 10%H 2O 2 method.Then Envision immunohistochemical stain with primary antibody Factor VIII-related antigen and vascular endothelial growth factor (VEGF). Results:All tissues showed Factor Ⅷ-related antigen and VEGF expression as well as the production were unaffected by bleaching.Compared with 10%H 2O 2,permanganate/oxalate can remove melanin granules more completely with much faster bleaching time,while both the antigenicities of FⅧ-Rag,VEGF and the tissue could be preserved. Conclusions:Potassium permanganate/oxalic acid melanin-bleaching technique can effectively bleach melanin in pigmented melanocytic lesions without significantly affecting FⅧ-RAg and VEGF immunolabelling.