目的   初步探讨生长因子受体结合蛋白14（GRB14）在肺腺癌患者预后中的具体作用机制。方法   通过TIMER数据库、UALCAN数据库及GEPIA数据库，探讨GRB14 mRNA在肺腺癌及正常肺组织中的表达。运用免疫组织化学通过组织芯片（75例肺腺癌患者和75例癌旁组织）检测其蛋白表达水平，收集国外肿瘤研究团队上传至TCGA数据库229例肺腺癌患者的临床数据，分析评估GRB14在肺腺癌患者的表达及其临床特征及生存预后之间的关系。应用TIMER数据库对GRB14肺腺癌患者进行免疫浸润分析。String数据库探讨GRB14与其他蛋白之间是否存在相互作用。结果  TIMER数据库分析显示，相比正常组织，GRB14 mRNA在多种实体肿瘤和肺腺癌组织中高表达（P＜0.05）。使用UALCAN数据库和GEPIA数据库以正常样本为对照组，肺腺癌患者的GRB14的表达均增加（P＜0.01）。免疫组织化学检测组织芯片结果显示，GRB14蛋白在肺腺癌的表达高于正常肺组织（肺腺癌6.07±1.01 vs 癌旁组织4.80±1.22；P＜0.01）。TCGA数据库分析显示，肺腺癌患者中GRB14高表达组和低表达组的中位总生存期分别为（41.59±5.20）月和（88.67±16.69）月；结合TCGA数据库绘制ROC曲线，发现GRB14的表达对肺腺癌患者具有一定的诊断价值。单因素回归分析结果显示，肿瘤分期（Ⅲ-Ⅳ）（P＜0.01）、肿瘤原发灶的情况（T3-4）（P＜0.01）、淋巴结转移（N1-3）（P＜0.01）和GRB14表达（P＜0.01）是影响肺腺癌中位总生存期的因素；Cox多因素回归分析显示，淋巴结转移（N1-3）（P＜0.05）和GRB14表达（P＜0.01）是影响肺腺癌中位总生存时间的因素。TIMER数据库分析显示，GRB14 mRNA 表达与巨噬细胞（r=-0.164，P＜0.01）、中性粒细胞（r=-0.175，P＜0.01）和树突状细胞（r=-0.148，P＜0.01）具有相关性。通过String数据库分析发现与GRB14相互作用的蛋白质包括EGFR、HRAS、FGFR1、INSR、CNGA1、COBLL1、LYPLAL1、TNKS2、TNKS、PRKCZ。结论  GRB14表达增加与肺腺癌患者预后不良相关。

       Objective  To assess the specific mechanism of growth factor receptor-bound protein 14（GRB14）in the prognosis of lung adenocarcinoma（LUAD）patients．Methods  The expression of GRB14 mRNA in LUAD and normal lung tissue was explored using TIMER database，UALCAN database，and GEPIA database．The expression of GRB14 protein was examined by immunohistochemistry using a tissue microarray．Then，the associations of GRB14 expression with clinicopathological features and clinical outcomes of LUAD were validated by analyzing TCGA database at the mRNA level and statistically evaluating the results．TIMER database was used to analyze immune infiltration of GRB14 in LUAD．Protein-protein interaction of GRB14 were analyzed using the String database．Results  Using the TIMER database，we found that GRB14 mRNA was highly expressed in various solid tumors and LUAD tissues compared to normal tissues（P＜0.05）．Comparing with the normal group，the expression of GRB14 was increased in LUAD（P＜0.01）via using UALCAN database and GEPIA database．The expression level of GRB14 protein in the LUAD tissues was significantly higher than that in the noncancerous LUAD tissues（LUAD［6.07±1.01］ vs benign，［4.80±1.22］；P＜0.01）in tissue microarray ．Median overall survival in the high and low GRB14 expression groups in LUAD was（41.59±5.2）and（88.67±16.69）months respectively．We plotted the ROC curves of 3-year survival rate and 5-year survival rate which again suggested that the model had good predictive performance．Univariate analysis revealed that individual cancer stages（Ⅲ-IV）（P＜0.01），tumor（T3-4）（P＜0.01），lymph node metastasis（N1-3）（P＜0.05）and GRB14 expression（P＜0.01）were risk factors affecting the median overall survival time of LUAD．According to Cox multiple regression analysis，we found that lymph node metastasis（N1-3）（P＜0.05）and GRB14 expression（P＜0.01）were  risk factors affecting the median overall survival time of LUAD．Using TIMER database analysis，the mRNA level of GRB14 was significantly correlated with macrophages（r=-0.164，P＜0.01），neutrophils（r=-0.175，P＜0.01）and dendritic cells（r=-0.148，P＜0.01）．Through analysis of the String database，it was found that proteins that interacted with GRB14 including EGFR，HRAS，FGFR1，INSR，CNGA1，COBLL1，LYPLAL1，TNKS2，TNKS，PRKCZ．Conclusions  The results of the present study suggest that GRB14 may efficiently predict poor survival in LUAD patients．

目的 探讨亚实性肺腺癌结节CT征象在预测侵袭风险中的价值。方法 选取经胸部CT检查发现并经病理证实为肺腺癌的亚实性结节共76个病灶,病灶分为侵袭前组与侵袭组2组,分析结节CT征象,行统计学分析,明确亚实性肺腺癌结节侵袭性的预测因素。结果 侵袭前组（包括原位腺癌、微浸润性腺癌）共31例;侵袭组[浸润性腺癌（IAC）]共45例。2组在最大平均直径、结节平均CT值、结节形状、分叶、毛刺、胸膜牵拉征、有无实性成分、肿瘤微血管CT成像征间比较差异有统计学意义（P<0.05）,而在空泡征、空气支气管征方面比较差异无统计学意义（P>0.05）。结节最大直径预测浸润性腺癌侵袭性界值为13.63 mm,敏感度、特异度分别为68.9%、96.8%,AUC为0.885;平均CT值预测IAC侵袭性界值为-528 HU,敏感度、特异度分别为84.4%、83.9%,AUC为0.867。回归分析显示病灶最大平均直径（OR=2.015、P=0.01）可以作为浸润性肺腺癌结节的独立预测因子。结论 亚实性肺腺癌结节侵袭前组和侵袭组在最大直径、平均CT值、形状、分叶、毛刺、微血管征存在差异;结节的最大平均直径是IAC的独立预测因子。

Objective To investigate the value of CT signs in predicting the risk of invasion of subsolid lung adenocarcinoma nodules．Methods A total of 76 subsolid pulmonary adenocarcinoma nodules detected by chest CT and pathologically confirmed were selected,and the lesions were divided into pre-invasion group and invasion group．CT signs of nodules were analyzed and statistical analysis was performed to determine the predictive factors of invasion of subsolid pulmonary adenocarcinoma nodules．Results There were 31 cases in the pre-invasion group（including adenocarcinoma in situ and minimally invasive adenocarcinoma）and 45 cases ininvasion group[invasive adenocarcinoma（IAC）]．There were significant differences in maximum mean diameter,mean CT value of nodule,nodule shape,lobule,burr,pleural stretch sign,solid component and microvascular CT imaging signs between the two groups（P<0.05）,but there were no significant differences in vacuole sign and air bronchial sign between the two groups（P>0.05）．The maximum diameter of nodules predicted the invasion boundary of invasive adenocarcinoma was 13.63 mm,the sensitivity and specificity were 68.9% and 96.8%,respectively,and the AUC was 0.885．The mean CT value predicted the invasive boundary of IAC was -528 HU,the sensitivity and specificity were 84.4% and 83.9%,respectively,and the AUC was 0.867．Regression analysis showed that the maximum mean lesion diameter（OR=2.015,P=0.01）was an independent predictor of infiltrating lung adenocarcinoma nodules．Conclusions There were differences in the maximum diameter,mean CT value,shape,lobed,burr and microvascular signs between the pre-invasion group and the invasion group．The maximum mean diameter of nodules was an independent predictor of IAC．

目的 使用TCGA数据库,探索DNA甲基化位点对肺腺癌的预后影响。方法 使用TCGA数据库,获取肺腺癌病人癌和癌旁组织甲基化表达数据、基因表达数据及临床数据;将人群分为探索组和验证组,使用LASSO在探索人群中筛选对肺腺癌预后有影响的甲基化位点;受试者工作特征曲线用于评估甲基化位点预测效果,并进一步在验证人群中验证。结果 在TCGA数据库中筛选出158个癌和癌旁组织差异表达且与所在基因mRNA表达显著相关的甲基化位点,经LASSO回归分析,cg19378330与肺腺癌预后相关。甲基化位点水平高于中位数的患者,归入高风险组,甲基化位点水平低于中位数的为低风险组。结果发现与低风险组相比,高风险组的死亡风险比低风险组增加了38%(OR=1.38,95% CI=1.16～2.69)。在探索阶段人群中其曲线下面积为0.80(95% CI=0.73～0.88),灵敏度为86.2%。验证人群中也表现出类似结果。结论 甲基化位点cg19378330与肺腺癌具有较显著的关联性,且可以对肺腺癌的风险进行有效的预测。

Objective Using the TCGA database to explore the prognostic effects of DNA methylation sites on lung adenocarcinoma. Methods TCGA database was used to collecting DNA methylation data, gene expression data and clinical data of lung adenocarcinoma patients. The population were divided into the exploratory group and the validation group. The LASSO regression analysis was used to screen the methylation sites associated with the prognosis of lung adenocarcinoma in the exploratory group. Receiver operating characteristic curve was used to evaluate the prediction effect of the model, and further verified in the validation population. Results A total of 158 methylation sites with differential expression and significant correlation with the mRNA expression of the corresponding gene were screened from the TCGA database. With LASSO regression analysis, the DNA methylation sites associated with prognosis of lung adenocarcinoma were cg19378330. Those patients with levels above the median methylation site were assigned to the high-risk group, while those with levels below the median methylation site were assigned to the low-risk group. Patients' death risk in the high-risk group were 38% higher than those in the low-risk group (OR=1.38, 95%CI=1.16-2.69). The area under the curve was 0.80 (95%CI=0.73-0.88) and the sensitivity was 86.2% in the exploratory stage population.Similar results were seen in the validation population. Conclusions The DNA methylation site cg19378330 was significantly associated withthe prognosisof lung adenocarcinoma, and could effectively predict the risk of lung adenocarcinoma.

目的 分析ABCC2基因表达水平与肺腺癌预后之间的关联性,并对其影响机制进行初步探索。 方法 采用TCGA数据库和HPA数据库对肺腺癌病人癌组织和癌旁组织基因表达数据进行差异性分析,单因素及多因素COX回归评估ABCC2与肺腺癌预后之间的关联性,GSEA用于探讨与ABCC2显著关联的信号通路。 结果 ABCC2在肺腺癌肿瘤组织中存在过表达现象,Kaplan-Meier生存分析曲线结果显示ABCC2基因过表达使肺腺癌病人的死亡风险显著升高(HR=1.46,95%CI=1.09~1.95; P=0.010)。单因素及多因素COX回归结果显示ABCC2基因过表达是肺腺癌病人不良预后的独立危险因素。GSEA结果显示ABCC2可能通过调节药物代谢从而对肺腺癌的发展进行调控。 结论 ABCC2基因过表达使肺腺癌病人的死亡风险显著升高,ABCC2可能是肺腺癌不良预后的潜在分子生物标志物。

Objective To estimate the association between ABCC2 mRNA expression and the prognosis of lung adenocarcinoma and explore the potential influencing mechanism.Methods Difference analysis was used to evaluate the gene expression in tumor tissues and adjacent normal tissues based on The Cancer Genome Atlas database and Human Protein Atlas database.Multivariate COX regression and Kaplan-Meier analysis were performed to evaluate the association between ABCC2 gene expression and the prognosis of lung adenocarcinoma.Gene-set enrichment analysis (GSEA) was performed to screen differentially enriched pathways associated with the ABCC2 high expression phenotype.Results ABCC2 was overexpressed in lung adenocarcinoma tumor tissues compared with adjacent normal tissues.Kaplan-Meier survival analysis showed a significant relationship between ABCC2 mRNA expression and lung adenocarcinoma prognosis (HR=1.16,95% CI=1.09-1.95; P=0.010).Univariate and multivariate Cox regression analysis showed that ABCC2 mRNA expression was an independent risk factor affecting the survival of patients with lung adenocarcinoma.The results of GSEA suggested that ABCC2 may influence the development of lung adenocarcinoma by regulating the metabolism of targeted drug the treatment.Conclusions ABCC2 overexpression can significantly increase the risk of death in patients with lung adenocarcinoma,ABCC2 may be a potential molecular marker for poor prognosis in lung adenocarcinoma.

目的 探讨Napsin A、TTF-1和CK7在恶性胸水细胞蜡块中的表达及对肺腺癌的诊断价值。方法 收集已确诊为恶性胸水且行Napsin A、TTF-1及CK7免疫组化标记,并收集患者的血清和胸水CEA,将患者按组织来源分为肺腺癌组和非肺腺癌组,比较Napsin A、TTF-1、CK7、血清CEA和胸水CEA在两组中的表达和浓度并计算它们的诊断价值。结果 相对于非肺腺癌组,肺腺癌组患者的Napsin A(83.9% vs 16.1%,P<0.001)、TTF-1(93.5% vs 6.5%,P<0.001)和CK7(98.1% vs 1.9%,P<0.001)阳性表达升高。Napsin A诊断肺腺癌的敏感度为83.9%,特异度为93.9%,TTF-1诊断肺腺癌的敏感度为93.5%,特异度为90.9%;CK7诊断肺腺癌的敏感度为98.1%,特异度为42.4%;明显高于传统肿瘤标志物血清CEA(诊断肺腺癌的敏感度为69.7%,特异度为58.5%)和胸水CEA(诊断肺腺癌的敏感度为69.0%,特异度为66.7%)。结论 Napsin A、TTF-1及CK7对于鉴别肺腺癌恶性胸水及其他肿瘤所致的恶性胸水具有较高的诊断价值。

Objective To investigate the expression of Napsin A, TTF-1 and CK7 in paraffin cells block of malignant pleural effusion and their diagnostic value for lung adenocarcinoma. Methods Immunohistochemistry of Napsin A, TTF-1, CK7, serum and pleural fluid CEA were collected from patients with malignant pleural effusion. The patients were divided into lung adenocarcinoma group and non-lung adenocarcinoma groups according to tissue source. The expression of Napsin A, TTF-1, CK7, CEA and the levels of serum and CEA in pleural fluid were compared and their diagnostic value was calculated. Results The positive expression of Napsin A (83.9% vs 16.1%, P<0.001), TTF-1 (93.5% vs 6.5%, P<0.001) and CK7 (98.1% vs 1.9%, P<0.001) in malignant pleural effusion caused by lung adenocarcinoma were higher compared to other tumors. The sensitivity and specificity of Napsin A were 83.9% and 93.9%,the sensitivity and specificity of TTF-1 were 93.5% and 90.9%, and those of CK7 were 98.1% and 42.4%; which were higher than those of serum CEA (69.7% and 58.5%) and pleural fluid CEA (69.0% and 66.7%) respectively. Conclusion Napsin A, TTF-1 and CK7 are of high value in the diagnosis of malignant pleural effusion caused by lung adenocarcinoma from other tumors.