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目的探讨SHH与PTCH在视网膜母细胞瘤(retinoblastoma,RB)组织中的表达及其在RB发生、发展中的意义。方法 17例手术切除的RB标本(RB组),17例因眼部外伤摘除的正常视网膜组织切片为对照组,2组分别采用实时PCR检测视网膜SHH mRNA与PTCH mRNA表达水平,采用Western blot法和免疫组织化学染色法检测视网膜SHH与PTCH蛋白表达水平,分析SHH mRNA和PTCH mRNA表达水平与RB临床病理特征间关系;采用Spearman等级相关分析SHH和PTCH蛋白在RB组织中表达的相关性。结果 RB组视网膜SHH mRNA和蛋白(2.86±0.57、5.19±1.61)及PTCH mRNA和蛋白(3.21±0.64、3.62±1.24)表达明显高于对照组(SHH mRNA和蛋白为1.02±0.13、0.56±0.06,PTCH mRNA和蛋白为1.14±0.17、0.74±0.15)(P<0.05);RB组SHH和PTCH蛋白阳性表达率(70.6%和58.8%)高于对照组(29.4%和23.5%)(P<0.05);RB组SHH mRNA和PTCH mRNA表达水平在肿瘤临床分期、淋巴结转移及分化程度上差异有统计学意义(P<0.05);RB组SHH表达与PTCH蛋白表达呈正相关(r=0.865,P=0.025)。结论 SHH与PTCH在RB中高表达,SHH信号通路被激活可在一定程度上反映RB的进展和预后。
Objective To investigate the expression of SHH and PTCH in retinoblastoma (RB) tissues and its significance in the development and progression of RB. Methods Seventeen RB specimens (RB group) and 17 normal retinal tissue sections removed by eye trauma were selected as control group. The expression of SHH mRNA and PTCH mRNA in retina were detected by real-time PCR in two groups. The expression of SHH and PTCH protein in retina was detected by immunohistochemical staining. The relationship between the expression of SHH mRNA and PTCH mRNA and clinicopathological features of RB was analyzed. Spearman rank correlation was used to analyze the correlation between SHH and PTCH protein expression in RB. Results The expressions of SHH mRNA and protein (2.86 ± 0.57,5.19 ± 1.61) and PTCH mRNA and protein (3.21 ± 0.64,3.62 ± 1.24) in RB group were significantly higher than those in control group (SHH mRNA and protein were 1.02 ± 0.13,0.56 ± 0.06 , PTCH mRNA and protein were 1.14 ± 0.17 and 0.74 ± 0.15 respectively) (P <0.05). The positive expression rates of SHH and PTCH in RB group were higher than those in control group (29.4% vs 23.5%, P < 0.05). The expressions of SHCH mRNA and PTCH mRNA in RB group were significantly different from those in the clinical stage, lymph node metastasis and differentiation (P <0.05). There was a positive correlation between SHH expression and PTCH protein expression in RB group (r = 0.865, P = 0.025). Conclusions SHH and PTCH are highly expressed in RB. The activation of SHH signaling pathway may reflect the progress and prognosis of RB to some extent.