视网膜母细胞瘤细胞系HXO-Rb_(44)抑癌基因的表达

The expression of tumor suppressor gene in the retinoblastoma cell line HXO Rb 44

目的检测视网膜母细胞瘤（ＲＢ）细胞系ＨＸＯＲｂ４４中抑癌基因Ｒｂ的表达状况，为进一步研究ＲＢ的发病机制，利用抑癌基因Ｒｂ进行ＲＢ的基因治疗提供实验依据。方法用ＳＰ免疫组织化学（ＩＨＣ）及流式细胞术（ＦＣＭ）间接免疫荧光法对Ｒｂ基因的表达进行定性、形态学定位及定量检测。结果ＩＨＣ法检测显示：正常视网膜组织中Ｒｂ蛋白呈阳性反应，位于细胞核呈棕黄色；ＨＸＯＲｂ４４中Ｒｂ蛋白表达阴性。ＦＣＭ检测显示：正常人淋巴细胞Ｒｂ蛋白表达量是ＨＸＯＲｂ４４细胞的３．７１倍，ＨＸＯＲｂ４４中Ｒｂ蛋白表达量极低。结论ＲＢ细胞系ＨＸＯＲｂ４４中存在野生型Ｒｂ基因的功能失活，Ｒｂ基因的突变失活与ＲＢ的发生密切相关。

ObjectiveTo detect the expression of the tumor suppressor gene Rb in the retinoblastoma (RB) cell line HXO Rb 44 for the further study of RB pathogenesis and providing experimental basis for the Rb gene therapy of RB in vivo.MethodsSP immunohistochemistry (IHC) was applied for the morphological observation-quality and localization detection and indirect immunofluorescence of flow cytometry (FCM) for the quantitative detection of the expression of Rb gene.ResultsIHC detection showed the positive expression of Rb protein with dark yellow staining in the nuclei of the normal retina tissues.Whereas most RB cells in HXO Rb 44 were negative staining of Rb protein.FCM quantitative detection in HXO Rb 44 showed much lower expression compared with normal tissues,and the positive expression of Rb gene in normal lymphocytes were 3.71 times as more as in RB cells in HXO Rb 44 .ConclusionOur results indicate that the loss of function of the wild type Rb gene was found in the RB cell line HXO Rb 44 and the mutation of wild type Rb gene was closely related to the RB formation.