The Application of Prognostic Model of Lysosomal Related Genes in Bladder Cancer
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摘要:
目的 探索基于溶酶体相关基因的预后模型在膀胱癌患者中应用的可行性。 方法 通过下载癌症基因组图谱(the cancer genome atlas program,TCGA)数据库中膀胱癌数据和基因表达综合数据库(gene expression omnibus,GEO)中GSE13507数据集。利于R语言通过差异分析、单因素比例风险模型(COX)回归分析筛选出TCGA数据库中与膀胱癌生存相关的差异表达的溶酶体相关基因,采用最小绝对值收敛和选择算子算法(Lasso)回归模型构建出预后模型。根据构建模型风险评分的中位值划分出高、低风险组。使用生存分析比较高、低风险2组患者的生存差异并在GEO数据集中进行验证。采用单因素及多因素Cox回归分析验证风险评分是否为影响膀胱癌患者预后的独立危险因素。受试者工作特征曲线用于评估预后模型预测的准确性。GO及KEGG富集分析用于探索高、低风险组差异基因的生物学功能及信号通路。免疫分析用于探索高、低风险组免疫功能差异。 结果 共筛选出44个差异表达的溶酶体相关基因,其中9个与预后相关基因用于预后模型构建,生存分析显示低风险组预后明显优于高风险组(P < 0.05),并在GEO数据库中得到验证。构建模型预测膀胱癌患者1 a、3 a、5 a生存的ROC曲线下面积(area under the curve,AUC)分别为0.696、0.717、0.738。独立预后分析提示构建模型为膀胱癌患者独立预后影响因素;GO富集分析提示高、低风险组差异基因主要参与细胞结构功能相关;KEGG富集分析提示差异基因主要富集于PI3K-Akt信号通路。免疫分析提示2组患者免疫细胞浸润情况及免疫功能具有明显差异(P < 0.05)。 结论 膀胱癌溶酶体相关基因风险模型能准确有效地预测膀胱癌患者预后。 Abstract:Objective To explore the feasibility of applying the prognosis model based on lysosomal related genes(LRGs) in patients with bladder cancer (BC). Methods Bladder cancer data were downloaded from The Cancer Genome Atlas (TCGA) program and dataset of GSE13507 were downloaded from Gene Expression Omnibus (GEO). Differentially expressed LRGs related to the survival of BC in the TCGA database were screened by differential analysis and single factor proportional hazards model (COX) regression analysis via R software. LASSO regression was used to construct a prognostic risk model. BC patients were divided into the high and low risk groups according to the median risk score. Survival analysis were used to compare the survival differences between the high-risk and low-risk groups of BC patients, and validate in GEO database. Univariate and multivariate cox regression analysis were used to verify whether the risk scores were an independent risk factor affecting the prognosis of BC patients. The receiver operating characteristic (ROC) curve was used to evaluate the accuracy of prognostic model predictions. GO and KEGG enrichment analysis were used to explore the biological functions and signaling pathways of differentially expressed genes between the high-risk and low-risk groups. Immunoassay was used to explore the differences in immune function between the high-risk and low-risk groups. Results A total of 44 differentially expressed lysosomal related genes were screened, of which 9 genes related to the prognosis were used to construct the prognosis model in this study. Survival analysis showed that the prognosis of the low-risk group was significantly better than that of the high-risk group (P < 0.05), which was verified in GEO database. The area under the ROC curve (AUC) of the BC prognosis risk scoring model to predict the 1-, 3- and 5-year survival of patients were 0.696, 0.717 and 0.738, respectively. Independent prognostic analysis showed that this prognostic risk model was an independent prognostic factor for BC patients. GO enrichment analysis indicated that the differential genes between the high-risk and low-risk groups were mainly involved in cell structure and function correlation. KEGG enrichment analysis suggested that the differential genes were mainly enriched in PI3K-Akt signaling pathway. Immunological analysis showed that there were significant differences in immune cell infiltration and immune function between the two groups. Conclusion Risk model of lysosomal related genes in BC can accurately and effectively predict the prognosis of BC patients. -
Key words:
- Bladder cancer /
- Lysosomal related genes /
- Prognostic model /
- Bioinformatics
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下颌磨牙的根管解剖非常复杂,根管治疗对牙髓专科医生是很大的挑战。我国人群的下颌磨牙根管解剖变异更为常见,尤其是下颌第二磨牙C形根管[1]及下颌第一磨牙额外牙根的变异[2]。本研究采用回顾性研究的方法分析云南昆明地区人群下颌第一和第二磨牙锥形束CT(Cone-beam computed tomography,CBCT)扫描数据,对于熟悉本地区人群下颌磨牙根管形态特征,提高治疗成功率具有非常重要的意义。
1. 资料与方法
1.1 研究对象
选取2018年10月至2018年12月于云南省第一人民医院口腔医学中心就诊行CBCT检查的所有患者总共403名,分析符合纳入标准的人群下颌第一和第二磨牙CBCT资料。
1.2 准入和排除标准[3]
牙根根尖孔完全闭合;无根尖周炎症、牙根吸收或折断影像;无金属或其他固定修复材料;年龄17~80岁。排除标准:已接受根管治疗;根尖孔未完全闭合;牙根吸收、折断或炎症;有金属或其他固定修复体。
1.3 扫描方法
所有患者采取安全保护措施,口腔放射科医师严格检查,仪器为RAYSCAN α-3D口腔数字化体层摄影及全景X射线机,电压电流90 kVp、4 mA,最小层面厚度300 μm、像素0.15 mm、扫描时间9~14 s、视野100 cm2(10 cm × 10 cm)。
1.4 图像分析
Xelis dental-CDViewer软件(1.0.5.2)分析下颌第一和第二磨牙轴状面(CEJ到根尖)、冠状面(颊舌向)、矢状面(近远中向)连续图像,记录牙根数量、根管数量和类型、C形根管在不同观察平面的类型。两名牙髓专科医师独立分析,有争议时与口腔放射科医师讨论决定。
1.5 统计学处理
SPSS软件(19.0)分析数据,卡方检验进行不同性别、牙位和年龄组间发生率的差异比较,P < 0.05为差异有统计学意义。
2. 结果
筛选分析403个CBCT图像,最终有358名患者1 229颗牙齿获得准入,其中男性183名,女性175名,下颌第一磨牙603颗、下颌第二磨牙626颗。
2.1 下颌第一和第二磨牙牙根及根管数量
本研究人群下颌第一磨牙总共603颗,其中大部分为典型2牙根(85.57%),少数有远舌根(14.43%),大部分为4根管(近远中各2根管,73.96%),19.73%为3根管、4.48%为5根管、1.49%为6根管、0.33%只有2根管,远舌根管发生率为14.43%,近中中根管发生率为3.15%(19/603),另有1个牙齿为C型根管。
下颌第二磨牙总共626颗,其中67.09%为2牙根,2.08%为3牙根,30.83%为融合牙根,8.94%为2根管,57.51%为3根管,2.72%为4根管,30.83%为C形根管,见表1、图1。
表 1 昆明地区人群下颌磨牙牙根及根管数量构成比[n(%)]Table 1. Number and distribution of roots and root canals of mandibular molars in a Kunming population [n(%)]牙位 牙根 根管 1 2 3 2 3 4 5 6 下颌第一磨牙 0(0) 516(85.57) 87(14.43) 2(0.33) 119(19.73) 446(73.96) 27(4.48) 9(1.49) 下颌第二磨牙 193(30.83) 420(67.09) 13(2.08) 56(8.94) 360(57.51) 17(2.72) − − 2.2 下颌第二磨牙C形根管发生率及类型特征
本研究人群626颗下颌第二磨牙中有193颗为融合牙根,C形根管平均发生率为30.83%,女性高于男性(34.95% > 26.81%,P < 0.05),左右侧无显著性差异(30.94%、30.72%,P > 0.05),各年龄组间无显著性差异(分别为47.37%、35.51%、23.08%、28.21%,P > 0.05),见表2。
表 2 昆明地区人群下颌第二磨牙C形根管发生率[n(%)]Table 2. Incidence of C-shaped canalsin mandibular second molars in a Kunming population [n(%)]项目 总数 C形根管
数量C形根管
发生率n(%)检验统计值 P值 性别 男 317 85 26.81 4.529 0.033 女 309 108 34.95 牙位 左 320 99 30.94 0.000 0.987 右 306 94 30.72 年龄
(岁)17~19 19 9 47.37 20~39 321 114 35.51 40~59 208 48 23.08 > 60 78 22 28.21 注:各年龄组比较:17~19岁与20~39岁比较,χ2 = 0.682,P = 0.409;17~19岁与40~59岁比较,χ2 = 2.576,P = 0.109;17~19岁与 > 60岁比较,χ2 = 2.469,P = 0.116;20~39岁与40~59岁比较,χ2=:3.562,P=0.059;20~39岁与 > 60岁比较,χ2 = 2.370,P = 0.124;40~59岁与 > 60岁比较,χ2 = 0.016,P = 0.899。 根据Fan’ s分类标准[4]对下颌第二磨牙C形根管在不同水平的类型进行分析,结果如下:根管口水平C1型为58.03%、C2型23.32%、C3(c)型6.74%、C3(d)型9.84%、C4型2.07%,根中水平C1型为8.29%、C2型38.86%、C3(c)型31.61%、C3(d)型17.10%、C4型4.14%,根尖水平C2型3.63%、C3(c)型12.95%、C3(d)型47.67%、C4型35.75%,未发现C5型,见表3。
表 3 昆明地区人群下颌第二磨牙C形根管构成比分析[n(%)]Table 3. Distribution of the C-shaped canals at different level in mandibular second molarsin a Kunming population [n(%)]部位 C1 C2 C3c C3d C4 合计 根管口 112(58.03) 45(23.32) 13(6.74) 19(9.84) 4(2.07) 193 根中部 16(8.29) 75(38.86) 61(31.61) 33(17.10) 8(4.14) 193 根尖3 mm 0(0.00) 7(3.63) 25(12.95) 92(47.67) 69(35.75) 193 3. 讨论
下颌第一磨牙常见的变异就是远舌根(radix entomolaris,RE),可独立,也可与其他牙根融合,其探查和疏通比较困难。Qiao等[5]分析四川地区人群1174颗下颌第一磨牙的CBCT资料,结果发现远舌根管发生率为22.15%,高于本研究人群,Al-Alawi H等[6]对884个沙特人的下颌第一磨牙形态进行研究,远舌根发生率仅为4.3%,低于本地区人群(14.43%)。笔者进一步对远舌根管形态进行分析,结果发现87.36%为Vertucci Ⅰ型,6.90%为Ⅱ型,5.75%为Ⅲ型。
下颌第一磨牙近中二根管间有时会有1个额外根管,称为近中中根管(mesial middle canal,MMC),可独立存在或融合,临床发生率为1%-15%[7]。Yang等[8]研究发现我国南方地区人群MMC发生率为9.03%,本研究人群603颗下颌第一磨牙绝大多数为4个根管,MMC发生率为3.15%。
下颌第二磨牙最常见的变异就是C形根管,有明显种族差异。Zheng等[9]研究发现中国西部人群下颌第二磨牙C形根管发生率为39%,与性别、年龄或牙位无关。本研究人群下颌第二磨牙C形根管发生率为30.83%,高于全球人群平均发生率(13.9%),女性较为常见,与年龄或牙位无关,这一点与Zheng等[9]研究结果有所区别。
Fan等[5]将C形根管分为5型:C1为连续C形、C2为分号形、C3c有3个独立根管、C3d有2个独立根管、C4为单根管、C5无根管影像。C形根管的形态从根管口到根尖孔移行过程中会有变化。Zheng等[9]发现从根管口到根尖孔C1和C2型根管逐渐减少,C3和C4型逐渐增多,与本研究结果一致。本研究人群C形根管在根管口绝大多数(81.35%)为C1和C2型,移行到中部变为C2和C3c型(70.47%),到根尖C3d和C4型又增多(83.42%),这意味着连续C形和分号形(C1和C2)很可能在根管中部或根尖分为2或3个根管,治疗时应特别注意。C1和C2型根管清理和成形比C3和C4型要困难得多,治疗时应加强化学预备。
昆明地区人群下颌第一磨牙远舌根管和第二磨牙C形根管的变异较常见,术前充分了解其根管形态特征,结合口腔手术显微镜和超声器械使用,可有效提升根管治疗成功率。
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1. 胡洁. 锥形束CT评估下颌第二磨牙根管治疗前后三维影像的临床价值. 现代医用影像学. 2022(11): 2120-2123 . 百度学术
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