目的:探讨肿瘤浸润免疫细胞(TIICs)在视网膜母细胞瘤(RB)患者预后中的作用,并构建基于多种免疫细胞的预后模型。方法:从GEO数据库获取RB样本和正常对照样本的基因表达谱及生存信息。(1) 使用CIBERSORT算法计算样本中不同免疫细胞的比例...目的:探讨肿瘤浸润免疫细胞(TIICs)在视网膜母细胞瘤(RB)患者预后中的作用,并构建基于多种免疫细胞的预后模型。方法:从GEO数据库获取RB样本和正常对照样本的基因表达谱及生存信息。(1) 使用CIBERSORT算法计算样本中不同免疫细胞的比例,进行免疫细胞浸润分析;(2) 进行Kaplan-Meier生存分析;(3) 通过LASSO回归、单因素和多因素Cox回归筛选与预后相关的免疫细胞,最终基于训练队列构建由3种免疫细胞(静息NK细胞、M0巨噬细胞和未活化CD4 T细胞)组成的预后模型;(4) 构建免疫细胞风险评分模型,并用Kaplan-Meier法评估模型;(5) 通过ROC曲线评估模型的预测能力。结果:在肿瘤样本中,树突状激活细胞、静息CD4记忆T细胞、调节性T细胞等免疫细胞活跃,而在正常样本中,初始B细胞和肥大细胞较为活跃。生存分析显示,低风险组患者的生存期显著长于高风险组。M0巨噬细胞和未活化CD4 T细胞与生存时间显著相关(P Objective: To investigate the role of tumor infiltrating immune cells (TIICs) in the prognosis of retinoblastoma (RB) patients, and to construct a prognostic model based on a variety of immune cells. Methods: The gene expression profiles and survival information of RB samples and normal control samples were obtained from GEO database. (1) The CIBERSORT algorithm was used to calculate the proportion of different immune cells in the sample, and the immune cell infiltration analysis was carried out;(2) Kaplan-Meier survival analysis;(3) Immune cells related to prognosis were screened by LASSO regression, univariate and multivariate Cox regression, and finally a prognostic model composed of three immune cells (resting NK cells, M0 macrophages and inactivated CD4 T cells) was constructed based on the training cohor;(4) Construct an immune cell risk scoring model and evaluate the model by Kaplan-Meier method;(5) The predictive ability of the model was evaluated by the ROC curve. Results: In tumor samples, immune cells such as dendritic activating cells, resting CD4 memory T cells, and regulatory T cells were active, while naïve B cells and mast cells were more active in normal samples. Survival analysis showed that patients in the low-risk group had significantly longer survival than those in the high-risk group. M0 macrophages and unactivated CD4 T cells were significantly correlated with survival time (P < 0.05). Conclusion: TIICs play an important role in the tumor microenvironment of RB, and specific immune cells (such as M0 macrophages, unactivated CD4 T cells, and resting NK cells) are significantly associated with patient survival. The prognosis model constructed in this study provides a new reference for clinical prognosis evaluation.展开更多
1 病例资料患儿男性,3岁,2020年12月因“发现右眼呈猫眼样反光”,就诊于我院,查眼底示“右眼视网膜母细胞瘤,IRC分期:E期”。2021年1月行右眼球摘除,术后病理示视网膜母细胞瘤,肿瘤侵犯视盘及筛板,穿透筛板达筛板后视神经内生长(距筛板...1 病例资料患儿男性,3岁,2020年12月因“发现右眼呈猫眼样反光”,就诊于我院,查眼底示“右眼视网膜母细胞瘤,IRC分期:E期”。2021年1月行右眼球摘除,术后病理示视网膜母细胞瘤,肿瘤侵犯视盘及筛板,穿透筛板达筛板后视神经内生长(距筛板后界约2.0mm),视神经切除断端未见明显肿瘤组织侵犯。免疫组化:NSE(+),Syn(+),CD56(+),Ki-67 index 70%。展开更多
文摘目的:探讨肿瘤浸润免疫细胞(TIICs)在视网膜母细胞瘤(RB)患者预后中的作用,并构建基于多种免疫细胞的预后模型。方法:从GEO数据库获取RB样本和正常对照样本的基因表达谱及生存信息。(1) 使用CIBERSORT算法计算样本中不同免疫细胞的比例,进行免疫细胞浸润分析;(2) 进行Kaplan-Meier生存分析;(3) 通过LASSO回归、单因素和多因素Cox回归筛选与预后相关的免疫细胞,最终基于训练队列构建由3种免疫细胞(静息NK细胞、M0巨噬细胞和未活化CD4 T细胞)组成的预后模型;(4) 构建免疫细胞风险评分模型,并用Kaplan-Meier法评估模型;(5) 通过ROC曲线评估模型的预测能力。结果:在肿瘤样本中,树突状激活细胞、静息CD4记忆T细胞、调节性T细胞等免疫细胞活跃,而在正常样本中,初始B细胞和肥大细胞较为活跃。生存分析显示,低风险组患者的生存期显著长于高风险组。M0巨噬细胞和未活化CD4 T细胞与生存时间显著相关(P Objective: To investigate the role of tumor infiltrating immune cells (TIICs) in the prognosis of retinoblastoma (RB) patients, and to construct a prognostic model based on a variety of immune cells. Methods: The gene expression profiles and survival information of RB samples and normal control samples were obtained from GEO database. (1) The CIBERSORT algorithm was used to calculate the proportion of different immune cells in the sample, and the immune cell infiltration analysis was carried out;(2) Kaplan-Meier survival analysis;(3) Immune cells related to prognosis were screened by LASSO regression, univariate and multivariate Cox regression, and finally a prognostic model composed of three immune cells (resting NK cells, M0 macrophages and inactivated CD4 T cells) was constructed based on the training cohor;(4) Construct an immune cell risk scoring model and evaluate the model by Kaplan-Meier method;(5) The predictive ability of the model was evaluated by the ROC curve. Results: In tumor samples, immune cells such as dendritic activating cells, resting CD4 memory T cells, and regulatory T cells were active, while naïve B cells and mast cells were more active in normal samples. Survival analysis showed that patients in the low-risk group had significantly longer survival than those in the high-risk group. M0 macrophages and unactivated CD4 T cells were significantly correlated with survival time (P < 0.05). Conclusion: TIICs play an important role in the tumor microenvironment of RB, and specific immune cells (such as M0 macrophages, unactivated CD4 T cells, and resting NK cells) are significantly associated with patient survival. The prognosis model constructed in this study provides a new reference for clinical prognosis evaluation.
文摘1 病例资料患儿男性,3岁,2020年12月因“发现右眼呈猫眼样反光”,就诊于我院,查眼底示“右眼视网膜母细胞瘤,IRC分期:E期”。2021年1月行右眼球摘除,术后病理示视网膜母细胞瘤,肿瘤侵犯视盘及筛板,穿透筛板达筛板后视神经内生长(距筛板后界约2.0mm),视神经切除断端未见明显肿瘤组织侵犯。免疫组化:NSE(+),Syn(+),CD56(+),Ki-67 index 70%。
文摘目的探讨长链非编码RNA(long non-coding RNA,LncRNA)核富集转录体1(nuclera-enriched autosomaltranscript,NEAT1)在视网膜母细胞瘤(retinoblastoma,Rb)患儿血清中表达,以及下调Rb细胞Y79中NEAT1对细胞生物学功能的影响。方法以2015年3月~2021年3月鄂东医疗集团黄石市中心医院诊疗的83例Rb患儿为研究对象,同期,在儿童保健中心选取健康儿童50例(对照组),实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测血清中NEAT1表达,分析Rb患儿和对照组血清NEAT1表达差异,以及不同临床指标Rb患者血清中NEAT1表达差异。培养Y79细胞并分为si-NEAT1组(转染NEAT1的干扰序列)、si-NC组(转染对照序列)和Ctl组(仅加入转染试剂),分别使用qRT-PCR,MTT,流式细胞术和Transwell检测NEAT1表达、细胞增殖、凋亡、迁移和侵袭情况。结果Rb患儿血清中NEAT1表达量(1.43±0.28)高于对照组(1.01±0.21),差异具有统计学意义(t=9.116,P<0.001);国际视网膜母细胞瘤分期(Intraocular International Retinoblastoma classificaton,IIRC)CDE期、低分化、视神经浸润和淋巴结转移的Rb患儿血清中NEAT1表达量明显高于AB期、中高分化、未发生视神经浸润和淋巴结转移的Rb患儿,差异具有统计学意义(t=2.190~3.693,均P<0.05);血清中NEAT1表达诊断Rb曲线下面积为0.882(95%CI:0.826~0.937),当NEAT1表达量取1.20时,灵敏度和特异度分别为80.00%和79.52%;相比于si-NC组(1.03±0.09)和Ctl组(1.02±0.15),si-NEAT1组细胞中NEAT1表达(0.35±0.06)明显降低,差异具有统计学意义(t=14.829,9.994,均P<0.001);si-NEAT1组24,48,72和96 h时吸光度(A值)明显低于si-NC组和Ctl组(tsi-NC=2.796~4.362,tCtl=2.641~5.555,均P<0.05),而细胞凋亡率相比于si-NC组和Ctl组明显升高,差异具有统计学意义(t=4.999,3.915,均P<0.05);与si-NC组和Ctl组比较,si-NEAT1组迁移细胞数(116.50±9.35 vs 132.00±7.32,134.00±7.95)和侵袭细胞数(96.33±8.94 vs 117.67±12.39,119.17±10.05)均降低,差异具有统计学意义(tsi-NC=3.196,3.421,tCtl=3.492,4.159,均P<0.05)。结论Rb患儿血清中NEAT1表达量升高,对Rb患儿具有一定的诊断价值,沉默Y79细胞中NEAT1表达可减少Rb细胞增殖、加速细胞凋亡,同时抑制细胞迁移和侵袭。
