lncRNA HOXA-AS2在视网膜母细胞瘤中的预后价值

Prognostic value of LncRNA HOXA-AS2 in retinoblastoma

目的探讨长链非编码RNA(lncRNA)HOXA-AS2在视网膜母细胞瘤组织中的预后价值。方法随机选取2018年5月就诊我科的3例视网膜母细胞瘤患儿的癌组织及同期就诊的3例正常视网膜样本,对两组间的组织样本进行lncRNA表达谱分析,筛选差异表达lncRNA。然后选择中挑选出HOXA-AS2,回顾性选取标本库中2012年1月至2017年12月就诊我科的60例视网膜母细胞瘤的手术标本进行PCR验证,根据HOXA-AS2表达水平进行分组,评估HOXA-AS2高表达组与HOXA-AS2低表达组的预后。最后预测HOXA-AS2的靶基因,并对靶基因数据集进行GO分析进行生物功能富集。结果对3例视网膜母细胞瘤和3例正常对照组的视网膜组织的lncRNA表达谱进行比较分析,共筛选出141个差异lncRNA,其中上调lncRNA共110个,下调lncRNA共31个。选择差异倍数最大的HOXA-AS2(logFC 4.54,P=3.94E-07)进一步验证。进一步搜集于2012年1月至2017年12月就诊我科的60例视网膜母细胞瘤的手术标本进行qRT-PCR验证,年龄2.5±1.8(0.6-6)岁,以HOXA-AS2表达中位值为截点进行分组,结果提示,HOXA-AS2高表达组的3年OS低于HOXA-AS2低表达组(51.2%比96.0%,P=0.006)。HOXA-AS2表达与不同性别、组织分型、肿瘤分期和视神经侵犯无关,与浸润有关,HOXA-AS2高表达组的脉络膜浸润率高于HOXA-AS2低表达组(35.5%比10.3%,P=0.021)。对视网膜母细胞瘤患儿的OS预后多因素分析,结果提示,较高的HOXA-AS2表达水平(OR=2.632,95%CI:1.228-5.641,P=0.013)是视网膜母细胞瘤患者预后(OS)的独立危险因素。运用Multi Experiment Matrix(MEM)在线数据库进行HOXA-AS2的靶基因预测,对HOXA-AS2的靶基因数据集进行GO分析,生物信息学分析结果提示主要富集于“早期胚胎发生后脑发育中前Hox基因的激活”的生物学过程中(FDR=5.60E-06)。结论视网膜母细胞瘤组织HOXA-AS2高表达可作为视网膜母细胞瘤不良预后的独立因素,有望成为视网膜母细胞瘤的预后标记物。

Objective To investigate the prognostic value of HOXA-AS2 in retinoblastoma.Methods Retinal tissues from randomly selected three patients with retinoblastoma and 3 normal subjects in our hospital during May 2018 were used in this study.The expression profiles of lncRNA were analyzed between the two groups,and the differential expression of lncRNA was screened.Then HOXA-AS2 was selected.Surgical specimens from 60 patients with retinoblastoma who were treated in our department from January 2012 to December 2017 were used for PCR verification.The study samples were divided to the HOXA-AS2 high-expression group and HOXA-AS2 low-expression group based on the expression level of HOXA-AS2,the prognosis were compared and evaluated between the two groups.Finally,the target gene of HOXA-AS2 was predicted and enriched by biological function(GO).Results By comparing the expression profiles of lncRNA in the retinal tissues of 3 cases of retinoblastoma and 3 cases of normal subjects,a total of 141 differential lncRNAs were identified,of which 110 were up-regulated and 31 were down-regulated.The HOXA-AS2 with the largest logFC was selected for further verification(logFC 4.54,P=3.94E-07).Another 60 surgical specimens of patients with retinoblastoma from January 2012 to December 2017 were used for qRT-PCR verification with an average age of 2.5±1.8(0.6~6)years,and were further grouped based on the median value of HOXA-AS2 expression level.The results showed that the OS of the HOXA-AS2 high expression group was lower than the low expression group at 3 years(51.2%vs.96.0%,P=0.006).The expression of HOXA-AS2 was not associated with gender,tissue type,tumor stage or optic nerve invasion,but related to infiltration.Choroidal infiltration rate in the HOXA-AS2 high expression group was higher than that the low expression group(35.5%vs.10.3%,P=0.021).Multivariate analysis showed higher HOXA-AS2 expression(OR=2.632,95%CI:1.228-5.641,P=0.013)was an independent risk factor for prognosis(OS)of retinoblastoma.Target gene prediction of HOXA-AS2 was performed using the Multi Experiment Matrix(MEM)online database and GO analysis was performed on the target gene data set of HOXA-AS2,the results indicated that it was mainly concentrated in the biological process of pre-Hox gene activation during brain development after early embryogenesis(FDR=5.60E-06).Conclusions The high expression of HOXA-AS2 in retinoblastoma tissues may be regarded as an independent prognostic factor for retinoblastoma,and may be a prognostic marker of retinoblastoma.