视网膜母细胞瘤的遗传发生机理

视网膜母细胞瘤(retinoblastoma,简称RB)是一种原发于视网膜组织的恶性眼内肿瘤。好发年龄为出生至7岁。多单眼患病,但也有30%～35%可双眼同时或先后患病。死亡率较高,一般在8%～65%之间,随肿瘤组     

视网膜母细胞瘤形成机理的研究进展

视网膜母细胞瘤(RB)系婴幼儿最常见的眼内恶性肿瘤,因其发病与遗传和环境因素有关,故一直被视为研究肿瘤遗传学及肿瘤形成机理的理想模式。近年来,不少遗传学和肿瘤学工作者对这方面进行了深入的研究,取得了一些新进展。本文从RB基因的研究进展、酯酶D的研究进展及其与RB的关系、限制性片断长度多态性技术在RB研究中的应用、RB基因及其基因产物的揭示、与RB有关的非随机染色体改变,以及RB形成理论及其实验证据等方面,简要进行综述。     

五种DNA肿瘤病毒在人视网膜母细胞瘤中检测不出

视网膜母细胞瘤(RB)是视网膜细胞缺乏功能性RB基因时诱发的小儿眼部肿瘤,近来研究表明:腺病毒、乳头状病毒,BK和JC多瘤病毒的转化蛋白能和RB基因产物结合,而且腺病毒12(Ad12)、JCK类人猿病毒SV40在啮齿动物中可诱发RB样肿瘤,鉴于以上发现,作者应用PCR技术对50例RB患者检测五种人DNA肿瘤病毒:Ad12、BKV、JCV、HPV16、HPV18,但未检测到这些病毒序列,试验结果表明:目前尚未有证据证实这些病毒是人RB的病因。人们认为当二个RB等位基因经过突变或丢失后才诱发肿瘤。遗传型RB,小儿遗传一个缺损性RB等位基因,在100百万正在发育的视网膜细胞中第二个等位基因的突变率很高足以导致多灶性双眼肿瘤发生,但只有     

视网膜母细胞瘤扫描电镜的组织形态学观察

从十六世纪中叶起,病理学家就开始对视网膜母细胞瘤(Retinablastoma)的组织病理学特征及其来源进行了研究。1864年Virehow(?)认为该肿瘤来自神经胶质细胞,故命名为神经胶质瘤。到1926年Verhoeff则认为它来自光感受细胞,命名为视网膜母细胞瘤,而延用至今。1981年有人通过组化染色方法进行     

视网膜母细胞瘤组织浸润程度临床相关危险因素研究

目的探讨视网膜母细胞瘤(Rb)对眼球组织浸润侵犯程度的相关临床危险因素。设计回顾性病例系列。研究对象经组织病理学确诊的Rb患者394例。方法收集1990-2005年北京同仁医院较完整的394例Rb患者的临床病理资料,术前观察主要临床体征,术后光镜下判读Rb眼球各组织浸润程度,通过logistic多元回归分析Rb眼球组织浸润程度的临床危险因素。主要指标Rb各临床体征与眼球组织浸润程度及两者之间的相关性。结果394例Rb中侵犯视盘315例(79．7％),侵犯筛板纤维203例(51．4％),肿瘤穿过筛板至球后视神经内102例(25．8％),侵犯脉络膜102例(25．8％),视神经切除断端受累者26例(6．6％)。统计分析显示:脉络膜组织受累与病程(P=0．032)、结膜充血(P=0．023)、眼球突出(P=0．026)及继发青光眼(P=0．027)有关;视神经切除断端浸润与结膜充血(P=0．008)、虹膜新生血管形成(P=0．033)、眼球突出(P=0．000)有关。结论本组Rb视神经侵犯率较高。病程长、结膜充血、眼球突出是眼球组织侵犯的主要因素。     

视网膜母细胞瘤中p16,pRb表达的研究

用免疫组化方法对２７例视网膜母细胞瘤中ｐ１６及ｐＲｂ的表达进行研究，结果发现４１％的病例不表达ｐ１６蛋白，５９％的病例不表达ｐＲｂ蛋白，两者均呈低表达的趋势，在一些病例中ｐ１６与ｐＲｂ有互换性缺失，提示ｐ１６及ｐＲｂ的视网膜母细胞瘤的发生中起重要作用。     

Hsp70在视网膜母细胞瘤组织芯片中的表达及其意义

目的检测Hsp70在视网膜母细胞瘤和正常视网膜组织中的表达,并讨论其意义。方法利用组织芯片,采用免疫组化SA法检测视网膜母细胞瘤和正常视网膜组织中Hsp70的表达。结果视网膜母细胞瘤组织芯片中Hsp70在胞浆有高表达(13/16),在正常视网膜组织中表达阳性率低(1/5),二者间存在显著性差异(P=0.0254<0.05)。结论Hsp70在视网膜母细胞瘤中高表达,它可能参与了视网膜母细胞瘤的发病。     

视网膜母细胞瘤铁蛋白表达的临床意义

目的:探讨视网膜母细胞瘤瘤组织中铁蛋白(FER)的分布、表达与肿瘤的临床及预后的关系以及作为肿瘤标记物的价值。方法:采用免疫组化方法观察视网膜母细胞瘤组织30例中FER的阳性分布。结果:FER阳性率96.6%(29/30),就诊年龄<2岁者,FER阳性范围以+++为主,占71.4%(5/7),阳性强度以++为主,占71.4%(5/7),就诊年龄>7岁者,阳性范围以+++为主,占83.3%(5/6),而就诊年龄在2～7岁之间者,阳性范围及强度均显示+为主(P<0.05);青光眼期患者,阳性范围以+++为主,占58.3%(7/12),眼内期++为主,占42.9%(3/7),眼外蔓延期+为主,占45.6%(5/11,P<0.05);分化型视网膜母细胞瘤阳性强度++为主,占85.7%(6/7),未分化型+为主,占65.2%(15/23,P<0.01)。结论:视网膜母细胞瘤中FER免疫组化的高表达可能是较好预后的一个临床病理指征。     

Fas系统在视网膜母细胞瘤的表达

目的 观察视网膜母细胞瘤（ＲＢ）中Ｆａｓ和Ｆａｓ配体（ＦａｓＬ）的表达特征。方法 应用免疫组化法检测３２例ＲＢ标本中Ｆａｓ和ＦａｓＬ的表达。结果 Ｆａｓ和ＦａｓＬ在ＲＢ中的阳性表达率分别为７０．６％和６５．８％,二者呈正相关（Ｐ〈０．００１）。分化程度低的和经过放疗的表达率明显增高（Ｐ〈０．０５或０．０１）。其表达率与视神经受浸润与否、眼内期或青光眼期无关（Ｐ〉０．０５）。结论 ＲＢ中Ｆａｓ及ＦａｓＬ高表达,分化程度越低表达越强。Ｆａｓ和ＦａｓＬ的表达可作为临床判断ＲＢ恶性程度的指标之一。     

Immunohistochemical localization of human papilloma virus in conjunctival neoplasias: a retrospective study

BACKGROUND: The extent of association of human papilloma virus (HPV) in human conjunctival neoplasias has been debated in studies originating from different parts of the world, but no substantial evidence has been generated on Indian subjects. This prompted us to carry out a retrospective study on conjunctival neoplasias diagnosed over the past 12 years. MATERIALS AND METHODS: Histopathological and immunohistochemical analysis of 65 specimens of ocular neoplasias and 30 normal controls diagnosed between 1991 and 2002 at a tertiary eye care hospital, was undertaken. Formalin-fixed, paraffin-embedded tissues were reviewed for confirming histopathological diagnosis, presence of koilocytosis and changes related to actinic keratosis. Immunohistochemical analysis was done using HPV-specific monoclonal antibodies. Clinicopathological correlation and the association of HPV antigen with the histopathological features were performed. RESULTS: Out of the 65 cases analyzed, 35 were papillomas and 30 were ocular surface squamous neoplasias (OSSN). The mean age was 48 years with a male preponderance. Histologically, koilocytosis was observed in 17.1% of papillomas and 36.6% of OSSN. Actinic keratosis was present in 33% of OSSN. Immunohistochemically 17.1% conjunctival papillomas stained positive for HPV antigen, all cases of OSSN were negative for HPV. There was no correlation between koilocytosis or actinic keratosis and the detection of HPV antigen. CONCLUSIONS: The association between HPV and conjunctival neoplasias is variable in different geographical areas and also depends on the methods of detection used. This study warrants the need for applying more advanced techniques at a molecular level to determine the possible etiology of HPV in conjunctival neoplasias among Asian-Indians.     

Development of genetically engineered tet HPV16-E6/E7 transduced human corneal epithelial clones having tight regulation of proliferation and normal differentiation

Lack of an optimal in vitro model of human corneal epithelial (HCE) cells is a major limitation in studying normal functions and gene regulations in HCE. Moreover, availability of a multi-layered HCE culture can reduce the usage of animals in the toxicity testing of consumer products. We have developed tetracycline-responsive human papilloma virus (HPV) 16-E6/E7 transduced HCE clones showing tight regulation of proliferation and normal differentiation. Expression of HPV16-E6/E7 mRNA and HPV16-E7 and keratin K3 proteins was examined by RNase protection assay and western blotting, respectively, in presence and absence (+/-) of Dox in identified clones. Localization of cornea-specific keratin k3 in +/- of Dox was evaluated by immunocytochemistry. The response of growth factors such as hepatocyte growth factor (HGF) and epidermal growth factor to the cellular proliferation in +/- of Dox in the newly identified clones was measured by cell counting. Cellular morphology, formation of multi-layered cultures at air-liquid interface and ultrastructural features were evaluated by light and transmission electron microscopy. The physical barrier established by the newly developed clones was determined by the transepithelial permeability to sodium fluorescein and transepithelial electrical resistance assays in the airlifted-stratified cultures.     

基质金属蛋白酶-2、基质金属蛋白酶-9和血管内皮生长因子在视网膜母细胞瘤中的表达

　　目的　观察基质金属蛋白酶（MMP）-2、MMP-9和血管内皮生长因子(VEGF)在视网膜母细胞瘤(RB)中的表达及其与RB分化程度和视神经浸润的关系。方法　经病理检查确诊的40例RB眼球石蜡标本纳入研究。按照RB细胞中有无菊形团排列以及RB肿瘤细胞是否浸润视神经分为分化型、未分化型以及视神经浸润、视神经无浸润组。其中,分化型15例,未分化型25例;视神经浸润13例,视神经无浸润27例。采用免疫组织化学方法检测其MMP-2、MMP-9和VEGF的表达,对比分析MMP-2、MMP-9和VEGF表达与肿瘤病理组织分型和视神经是否侵润之间的关系。结果　40例RB中,MMP-2、MMP-9和VEGF的阳性表达率分别为52.5%、57.5% 和72.5% 。未分化型RB中MMP-2、MMP-9和VEGF的阳性表达均高于分化型RB中MMP-2、MMP-9和VEGF的阳性表达,差异有统计学意义(χ²=9.037,9.253,8.095;P＜0.05);有视神经浸润的RB中MMP-2、MMP-9和VEGF的阳性水平均高于无视神经浸润RB中MMP-2、MMP-9和VEGF的阳性水平,差异有统计学意义(χ²=11.045,10.243,8.956;P＜0.05)。RB中MMP-2、MMP-9和VEGF的表达呈正相关关系(r=0.126,0.314;P＜0.05)。结论　RB细胞内有MMP-2、MMP-9和VEGF的表达。MMP-2、MMP-9表达与VEGF表达存在相关性。MMP-2、MMP-9和VEGF的表达水平与视神经浸润存在密切的关系。       

HSP60在人视网膜母细胞瘤中表达的研究

目的 研究热休克蛋白60(heat shock protein 60,HSP60)在未接受治疗的视网膜母细胞瘤(retinoblastoma,Rb)患者的视网膜内原发肿瘤细胞和脉络膜、视神经等侵袭部位的肿瘤细胞中的表达,初步分析HSP60在眼球不同部位Rb细胞的表达特点及其可能与Rb细胞凋亡的关系.方法 收集22例(22眼)未接受治疗的Rb患者所摘除眼球的石蜡切片.用免疫组织化学方法测定HSP60、HSP10和Caspase 3在视网膜内和脉络膜、视神经等侵袭部位的表达.结果 22例患者手术后病理检查眼内均见肿瘤细胞.侵袭部位见肿瘤细胞的患者21例,其中肿瘤单独侵犯视神经7例(33.3％),肿瘤单独侵犯脉络膜5例(23.8％),肿瘤同时侵犯视神经和脉络膜9例(42.9％).脉络膜与视神经部位肿瘤细胞HSP60、HSP10的表达均比眼内肿瘤细胞高,差异均有统计学意义(均为P＜0.05).脉络膜与视神经部位肿瘤细胞Caspase 3的表达比眼内肿瘤细胞低,差异均有统计学意义(均为P=0.000).结论 Rb肿瘤细胞在从视网膜内向外侵袭的过程中,可能通过升高HSP60、降低Caspase 3来促进侵袭部位肿瘤细胞的存活.但是HSP60在侵袭部位的升高是肿瘤细胞向外侵袭的原因还是结果,以及是否会进一步增强肿瘤细胞向远处转移等还需进一步研究.     

Expression of multidrug resistance proteins in retinoblastoma

AIM: To elucidate the mechanism of multidrug resistance in retinoblastoma, and to acquire more insights into in vivo drug resistance. METHODS: Three anticancer drug resistant Y79 human RB cells were generated against vincristine, etoposide or carboplatin, which are used for conventional chemotherapy in RB. Primary cultures from enucleated eyes after chemotherapy (PCNC) were also prepared. Their chemosensitivity to chemotherapeutic agents (vincristine, etoposide and carboplatin) were measured using MTT assay. Western blot analysis was performed to evaluate the expression of p53, Bcl-2 and various multidrug resistant proteins in retinoblastoma cells. RESULTS: Following exposure to chemotherapeutic drugs, PCNC showed less sensitivity to drugs. No significant changes observed in the p53 expression, whereas Bcl-2 expression was found to be increased in the drug resistant cells as well as in PCNC. Increased expression of P-glycoprotein (P-gp) was observed in drug resistant Y79 cells; however there was no significant change in the expression of P-gp found between primary cultures of primarily enucleated eyes and PCNC. Multidrug resistance protein 1 (Mrp-1) expression was found to be elevated in the drug resistant Y79 cells as well as in PCNC. No significant change in the expression of lung resistance associated protein (Lrp) was observed in the drug resistant Y79 cells as well as in PCNC. CONCLUSION: Our results suggest that multidrug resistant proteins are intrinsically present in retinoblastoma which causes treatment failure in managing retinoblastoma with chemotherapy.     

眼发育调控基因Pax6在视网膜母细胞瘤中表达的初步研究

目的 探讨眼发育调控基因Pax6在人视网膜母细胞瘤中的表达情况.设计 实验性研究.研究对象 新鲜视网膜母细胞瘤肿瘤组织6例,正常视网膜组织6例.视网膜母细胞瘤细胞株2例.方法 采用逆转录酶链反应(RT-PCR)、Western蛋白印迹方法检测视网膜母细胞瘤肿瘤组织中Pax6 mRNA和Pax6蛋白的表达,以正常视网膜组织作为对照.同时检测SO-Rb50、Y79视网膜母细胞瘤细胞株中Pax6 mRNA和Pax6蛋白的表达情况.主要指标 视网膜母细胞瘤新鲜肿瘤组织及细胞株中Pax6基因的表达.结果 RT-PCR和Western蛋白印迹检测显示,在6例视网膜母细胞瘤肿瘤组织及两种视网膜母细胞瘤细胞株中均可见Pax6 mRNA及Pax6蛋白表达.与正常视网膜组织相比较,视网膜母细胞瘤肿瘤组织中Pax6 mRNA及Pax6蛋白表达水平增高(P均<0.01).结论 Pax6基因的异常表达可能参与并导致人眼视网膜母细胞瘤的发生和发展.     

E26转录因子-1在视网膜母细胞瘤中的表达及意义

目的探讨E26转录因子-1（Ets-1）在视网膜母细胞瘤（Rb）中的表达及其与肿瘤的血管化、浸润、转移的关系。方法利用原位杂交和免疫组织化学法分别检测Ets-1mRNA和蛋白在40例Rb中的表达。并以免疫组织化学法检测肿瘤中CD34的表达，以此计算肿瘤微血管密度的值。结果82．5％（33／40）的Rb中Ets-1mRNA呈阳性表达，Ets-1蛋白的阳性表达率为65％（26／40）。Ets-1主要表达于癌细胞和血管内皮细胞。Ets-1高表达组的微血管密度明显高于Ets-1低表达组（P〈0．01）。其表达与组织学分级和预后有关。结论Els-1可促进Rb的新生血管形成，与肿瘤的浸润和转移有关。     

棘阿米巴性角膜炎的发病相关因素

由于角膜接触镜的日益广泛使用,棘阿米巴性角膜炎在世界范围内呈上升趋势。本文就棘阿米巴性角膜炎发病相关因素包括人口学特征（性别、年龄、职业）、季节倾向和地理分布特征以及易感危险因素（角膜接触镜佩戴、眼部创伤、眼部手术、并存眼部疾病或混合感染）的研究进展进行综述。（国际眼科纵览,2020, 45：393-396）