综合治疗前后视网膜母细胞瘤微血管密度和血管内皮生长因子表达研究

Microvessel density and expression of vascular endothelial growth factor in retinoblastoma before and after comprehensive treatment

目的 观察视网膜母细胞瘤（RB）未行综合治疗和行综合治疗后微血管密度（MVD）、血管内皮生长因子（VEGF）表达以及与RB肿瘤视神经浸润和综合治疗后复发的关系.方法 61例RB石蜡标本纳入研究.分为未行综合治疗直接摘除眼球（未治疗）组、综合治疗前计划眼球摘除（计划眼摘）组、综合治疗后肿瘤复发（复发）组,3组分别为52、6、3例.其中,未治疗组中,视神经未侵润33例,已侵润19例.应用CD34单克隆抗体、VEGF单克隆抗体、链霉菌抗生物素蛋白-过氧化物酶连结法（SP）免疫组织化学法,对61例石蜡标本行MVD、VEGF表达测定;免疫组织化学和实时定量聚合酶链反应方法 检测计划眼摘组和复发组VEGF的表达,观察MVD、VEGF表达.结果 未治疗组RB浸润视神经者其MVD多于未浸润视神经者,二者比较,差异有统计学意义（t=-2.468 5,P=0.017）.RB浸润者视神经VEGF表达高于未浸润视神经者,二者比较,差异有统计学意义（χ2=8.416 6,P=0.032 8）.计划眼摘组,在全身化学治疗过程中,肿瘤组织微血管消退,MVD降低,部分血管闭塞,玻璃样变.复发组肿瘤组织中可见大量新生微血管,MVD增加.计划眼摘组的VEGF表达低于复发组VEGF表达.结论 未行综合治疗与行综合治疗者之间RB VEGF阳性表达差异无统计学意义;未行综合治疗者RB MVD增加、VEGF阳性表达均与肿瘤视神经浸润有关;行综合治疗后RB复发可能与MVD增加有关.

Objective To observe the microvessel density （MVD）and expression of vascular endothelial growth factor （VEGF） in retinoblastoma（RB）before and after comprehensive treatment and explore its clinical correlations with tumor infiltration and recurrence. Methods Sixty-one paraffin specimens of RB were divided into enucleation without comprehensive treatment group （untreated group,52cases）, planned enucleation before comprehensive treatment group （planned enucleation group, six cases）and tumor recurrence after comprehensive treatment group（recurrence group,three cases）. There were optic nerve invasion in 19 cases,no optic nerve infiltration in 33 cases. The MVD and VEGF expressions of 61paraffin specimens were detected by streptavidin biotin-peroxidase （SP） immunohistochemistry with monoclonal antibody of CD34 and VEGF. Real time PCR was performed for the VEGF expression of planned enucleation group and recurrence group. Results In the untreated group, the MVD and VEGF expression of optic nerve infiltration cases were significantly higher than those of cases without optic nerve infiltration（t= -2. 4685, P=0. 017;χ2 =8. 416 6,P=0.032 8）. Tumor microvessel regression, decreased MVD, occlusion and hyaline changes of blood vessels were observed in the planned enucleation group in the course of systemic chemotherapy. Many neovascularized capillaries and increased MVD were observed in tumor tissues of the recurrence group. The VEGF expression of planned enucleation group was lower than that in the recurrence group. Conclusions There was no significant difference on VEGF expression in RB between with and without comprehensive treatment. The increasing MVD and VEGF expression of cases without comprehensive treatment were related to the optic nerve infiltration. And the increasing MVD may also play an important role in RB recurrence after comprehensive treatment.