Identification of key genes and biological pathways in lung adenocarcinoma by integrated bioinformatics analysis

BACKGROUND The objectives of this study were to identify hub genes and biological pathways involved in lung adenocarcinoma(LUAD)via bioinformatics analysis,and investigate potential therapeutic targets.AIM To determine reliable prognostic biomarkers for early diagnosis and treatment of LUAD.METHODS To identify potential therapeutic targets for LUAD,two microarray datasets derived from the Gene Expression Omnibus(GEO)database were analyzed,GSE3116959 and GSE118370.Differentially expressed genes(DEGs)in LUAD and normal tissues were identified using the GEO2R tool.The Hiplot database was then used to generate a volcanic map of the DEGs.Weighted gene co-expression network analysis was conducted to cluster the genes in GSE116959 and GSE-118370 into different modules,and identify immune genes shared between them.A protein-protein interaction network was established using the Search Tool for the Retrieval of Interacting Genes database,then the CytoNCA and CytoHubba components of Cytoscape software were used to visualize the genes.Hub genes with high scores and co-expression were identified,and the Database for Annotation,Visualization and Integrated Discovery was used to perform enrichment analysis of these genes.The diagnostic and prognostic values of the hub genes were calculated using receiver operating characteristic curves and Kaplan-Meier survival analysis,and gene-set enrichment analysis was conducted.The University of Alabama at Birmingham Cancer data analysis portal was used to analyze relationships between the hub genes and normal specimens,as well as their expression during tumor progression.Lastly,validation of protein expression was conducted on the identified hub genes via the Human Protein Atlas database.RESULTS Three hub genes with high connectivity were identified;cellular retinoic acid binding protein 2(CRABP2),matrix metallopeptidase 12(MMP12),and DNA topoisomerase II alpha(TOP2A).High expression of these genes was associated with a poor LUAD prognosis,and the genes exhibited high diagnostic value.CONCLUSION Expression levels of CRABP2,MMP12,and TOP2A in LUAD were higher than those in normal lung tissue.This observation has diagnostic value,and is linked to poor LUAD prognosis.These genes may be biomarkers and therapeutic targets in LUAD,but further research is warranted to investigate their usefulness in these respects.

Kinesin KIF4A is associated with chemotherapeutic drug resistance by regulating intracellular trafficking of lung resistance-related protein

Multidrug resistance （MDR） is the major impediment to cancer chemotherapy. The expression of lung resistance-related protein （LRP）, a non-ATP-binding cassette （ABC） transporter, is high in tumor cells, resulting in their resistance to a variety of cytotoxic drugs. However, the function of LRP in tumor drug resistance is not yet explicit. Our previous studies had shown that Kinesin KIF4A was overexpressed in cisplatin （DDP）-resistant human lung adenocarcinoma cells （A549/DDP cells） compared with A549 cells. The expression of KIF4A in A549 or A549/DDP cells significantly affects cisplatin resistance but the detailed mechanisms remain unclear. Here, we performed co-immunoprecipitation experiments to show that the tail domain of KIF4A interacted with the N-terminal of LRP. Immunofluorescence images showed that both the ability of binding to LRP and the motility of KIF4A were essential for the dispersed cytoplasm distribution of LRP. Altogether, our results shed light on a potential mechanism in that motor protein KIF4A promotes drug resistance of lung adenocarcinoma cells through transporting LRP-based vaults along microtubules towards the cell membrane. Thus KIF4A might be a cisplatin resistance-associated protein and serves as a potential target for chemotherapeutic drug resistance in lung cancer.

Expression of Livin in tissues of lung cancer and its correlation with the expression of caspase-3

Objective： To study the expressions of two isoforms of Livin in tissues of lung cancer and their relations to histological types and chemotherapy, and to study their correlations to the expression of caspase-3 as well. Methods： Expressions of Livin isoforms a, 13 and caspase-3 were detected by reverse transcription polymerase chain reaction （RT-PCR） assay in lung cancer tissues as well as in controls. Results： Livin isoforms a and ~ were expressed in 12 of 27, and 19 of 27 lung cancer tissues respectively, much more than those in lung para-cancereus [both were （0/6）] or benign disease lung tissues （0/12, 1/12; P 〈 0.01 and P 〈 0.01 ）. Moreover, they were detected in 7/14, 9/14 lung adenocarcinomas and 4/12, 9/12 squamocallular and large call carcinomas, respectively, and both showed expressions in one small cell carcinoma. The levels of these two isoforms in lung cancer were significantly higher than those in controls by Gel imaging system （P 〈 0.05 and P 〈 0.05）, the former was higher in adenocarcinoma than that in squamocellular carcinoma （P 〈 0.05）, while the latter was the same in both （P 〉 0.05）. Meanwhile, the levels of caspase-3 in lung cancer were significantly lower than those in controls, and it was suggested to be negatively associated with either each of two isoforms or their sum （P 〈 0.05, P 〈 0.01 and P 〈 0.01）. Two isoforms of Livin expression seemed to increase＇after chemotherapy but not related to clinical stages （P 〉 0.05）. Conclusion： Two isoforms of Livin are differently expressed in different histological types of lung cancer and may contribute to corresponding cancerous development; the levels of Livin are negatively associated with those of caspase-3, this may be due to the fact that Livin could resist against apoptosis; high expression of Livin seems to be related to chemotherapy but not clinical stages.

Expression of Coxsackievirus and Adenovirus Receptor in Human Lung Cancer： Possible Clinical Significance

OBJECTIVE To explore the relationship between CAR and the development of human lung cancer, as well as to provide the basis for the clinical treatment of lung cancer using an adenovirus vector-based gene therapy. METHODS CAR expression was assessed immunohisto- chemically in tumoral, paraneoplastic and normal samples from 112 lung cancer patients. At the same time, the mRNA and protein expression of CAR in 32 cases were determined by RT-PCR and Western blot. The relationship between CAR expression and clinicopathologic parameters was statistically analyzed. RESULTS There was no expression of CAR in normal lung tissue but a little in paraneoplastic tissue. The positive rate was 43% in squamous cell carcinoma, and 70% in adenocarcinoma. Both were much significantly higher than that in paraneoplastic tissue. The CAR expression level in adenocarcinoma was higher than that in squamous cell cancer, mRNA expression by RT-PCR and protein expression by Western blot were consistent with immunohistochemistry results. CONCLUSION CAR is overexpressed in human lung cancer, especially in adenocarcinoma. This data offer the reliable basis for adenovirus-mediated gene therapy of lung cancer; more important, CAR may take part in the formation or development of lung cancer; this may be exploitable for the development of antibody-directed therapy in human lung cancer.

Cetuximab Combination with Chemotherapy in Advanced Non-Small Cell Lung Cancer

Objective： To observe the efficacy and safety of cetuximab combined with chemotherapy in advanced non-small-cell lung cancer （NSCLC）, and to investigate the association of status of K-RAS gene mutation and epidermal growth factor receptor （EGFR） genotype with clinical outcome. Methods： Between Jan. 2006 and Sep. 2009, nineteen patients with advanced NSCLC received cetuximab （〉4 weeks） combined with chemotherapy in Department of Thoracic Oncology at Beijing Cancer Hospital. Response, survival and toxicity were retrospectively assessed, epidermal growth factor receptor （EGFR） protein expression was evaluated by ELISA Kit. The status of K-RAS gene mutation was tested by PCR-RFLP and EGFR gene amplification was measured by EGFR fluorescence in situ hybridization （FISH）. Results： Partial response（PR） was observed in 26.3%（5/19） of the patients and stable disease（SD） in 52.6%（10/19）. Median progression free survival（PFS） was 6 months （95% CI： 3.6-8.4）. Median overall survival （MST） and 1-year survival rate（SR） were 10.6 months （95% CI： 6.6-14.6） and 47.6%, respectively. Mild or moderate skin rash was the most common toxicity related with cetuximab. K-RAS gene mutation, EGFR protein level and amplification have little correlation with prognosis. Conclusion： Cetuximab combined with chemotherapy was tolerable and the skin rash related with cetuximab was mild to moderate. Cetuximab may prolong survival of the patients who failed to previous chemotherapy.

CD8 T cell response in a phase I study of therapeutic vaccination of advanced NSCLC with allogeneic tumor cells secreting endoplasmic reticulum-chaperone gp96-Ig-peptide complexes

Antigen containing, allogeneic cells secreting the genetically modified protein and peptide-chaperone gp96-Ig cross, prime and expand antigen specific CD8 T cells with therapeutic antitumor activity in mice. In a first in man phase I study, we now report the results of therapeutic vaccination of non-small cell lung cancer (NSCLC) patients with an established, allogeneic non-small cell lung adenocarcinoma cell line secreting gp96-Ig. Advanced NSCLC-patients stage IIIB or IV of any histological subtype were enrolled and treated with up to 36 vaccinations over the course of 18 weeks. Primary endpoint was safety, secondary endpoints tumor response and overall survival. Measurement of tumor antigen specific CD8 CTL responses is precluded by the lack of known NSCLC associated antigens. Therefore, we measured patient CD8 T cell-IFN-γ responses to allo-antigens of the vaccine cells as surrogate for tumor antigen specific CD8 CTL. In 7 of 18 treated patients tumor growth was stabilized, however none of the 18 patients had an objective tumor response by RECIST criteria. Of 15 patients evaluable for immune response, 11 responded to vaccination with more than twofold increase in CD8-IFN-γ frequency above baseline. These patients had a median survival time of 16.5 months. Four patients who had no CD8 response above base line had survival times from 2.1 to 6.7 months. Our data are consistent with the concept that generation of CD8 CTL by therapeutic vaccination may delay tumor growth and progression and mediate prolonged survival even in the absence of objective tumor responses. Further studies will be required to test this concept and promising result.

A Study on the expression of erbB4/HER4 in non-small cell lung cancer

Objective: To test the expression of HER4 in non-small cell lung cancer (NSCLC) and elucidate the relationship between its over-expression and the clinical pathology of NSCLC. Methods: 70 cases of paraffin-embedded tissues from informative NSCLC were tested for the expression of HER4 by means of immunohistochemical assay. Results: HER4 were overexpressed in NSCLC in 91.4%. The overexpression of HER4 correlated only with the lymph node metastasis, TNM staging and survival after operation. Conclusion: ErbB4 is one of the genes to regulate the growth of NSCLC in advanced stages and artificial interference of the overexpression of HER4 in NSCLC might be a good way for the treatment of NSCLC in advanced stages.

Effects of chimeric intron on the epithelial-mesenchymal transformation of non-small cell lung cancer PC-9

Objective:To investigate the effects of Intron on the EMT capability of non-small cell lung cancer cell line PC-9.Methods:Firstly,using the psiCHECK-2 plasmid as a basic framework to construct the recombinant plasmid of psiCHECK-2-Intron dual-luciferase reporter gene;secondly,the psiCHECK-2-Intron and psiCHECK-2 were transfected into PC-9 cells respectively.The migration and invasion abilities of PC-9 cells were analyzed by Matrigel assay.The expression changes of EMT related hallmarks,including N-cadherin,β-catenin and snail,were detected by qRT-PCR and Western Blotting.Results:Compared with the control group,the migration and invasion abilities of PC-9 cells in Intron group significantly decreased(p<0.001).The expression of N-cadherin,β-catenin and snail also down-regulated(p<0.001).Conclusion:The introns could inhibit the EMT of PC-9 cells.

非小细胞肺癌组织EMSY、PIDD表达与同源重组修复基因的相关性及其临床意义

目的研究非小细胞肺癌(NSCLC)组织中EMSY转录抑制因子(EMSY)、p53诱导的死亡结构域蛋白1(PIDD)表达与同源重组修复基因表达的相关性及临床意义。方法选择2022年1月—2023年4月河北北方学院附属第一医院呼吸与危重症医学科诊治NSCLC患者80例。采用实时荧光定量PCR检测癌组织及癌旁组织中EMSY、PIDD表达与同源重组修复基因人乳腺癌易感基因1(BRCA1)、切除修复交叉互补基因1(ERCC1),核糖核苷酸还原酶亚单位1(RRM1)表达。Pearson相关分析EMSY、PIDD表达与同源重组修复基因表达的相关性;分析EMSY、PIDD表达与NSCLC临床病理相关参数的关系及在NSCLC诊断中的价值。结果NSCLC癌组织中EMSY、PIDD、BRCA1、ERCC1、RRM1 mRNA相对表达量均高于癌旁组织(t/P=30.176/<0.001,27.821/<0.001,25.075/<0.001,16.680/<0.001,25.610/<0.001)。NSCLC癌组织中EMSY、PIDD mRNA与BRCA1、ERCC1、RRM1 mRNA表达均呈正相关(r/P=0.654/<0.001,0.712/<0.001,0.584/<0.001;0.724/<0.001,0.661/<0.001,0.563/<0.001)。低分化程度、有淋巴结转移及TNM分期Ⅲ期NSCLC癌组织EMSY、PIDD mRNA表达分别显著高于高中分化程度、无淋巴结转移及TNM分期Ⅰ~Ⅱ期NSCLC癌组织(t/P=13.693/<0.001,13.380/<0.001,12.197/<0.001;10.289/<0.001,11.130/<0.001,9.405/<0.001)。EMSY、PIDD mRNA及两项联合诊断NSCLC的AUC分别为0.834、0.802、0.906,两项联合诊断的AUC大于单一指标(Z=4.751、5.257,P均<0.001)。结论EMSY、PIDD在NSCLC癌组织中表达升高,与同源重组修复基因表达及不良临床病理特征有关,两者联合有助于NSCLC的诊断。

ERCC1、K-ras、TP-73在替雷利珠单抗联合TP化疗方案治疗非小细胞肺癌中的评估价值

目的研究探讨核苷酸切除修复交叉互补基因1(ERCC1)、Kirsten-Rous肉瘤病毒蛋白(K-ras)、肿瘤蛋白P73(TP73)在替雷利珠单抗结合紫杉醇+顺铂(TP)化疗方案治疗NSCLC中的评估价值。方法选取2020年1月至2021年12月本院收治的126例NSCLC肺癌患者为研究对象,按随机抽签法分为对照组、观察组,各63例。对照组以TP化疗方案治疗,观察组增加替雷利珠单抗治疗。评估组间临床疗效、肿瘤标记蛋白、免疫指标、生存周期、不良反应。结果观察组患者的客观缓解率为69.84%(44/63)高于对照组患者为52.38%(33/63),观察组疾病控制率为82.54%(52/63),高于对照组患者为66.67%(42/63)(P<0.05)。化疗1周期、化疗3周期、化疗6周期时,观察组ERCC1、K-ras、TP-73水平均低于对照组(P<0.05)。治疗后观察组免疫功能补体C3、补体C4、CD40细胞低于对照组,NK细胞高于对照组(P<0.05)。观察组患者的TTP、PFS、总生存期均高于对照组(P<0.05)。观察组不良反应发生率为19.05%(12/63),对照组为12.70%(8/63),组间比较差异无统计学意义(P>0.05)。结论替雷利珠单抗联合TP化疗方案治疗肺癌有良好的治疗效果,能够改善患者免疫功能,延长患者生存周期,治疗安全性较好,且ERCC1、K-ras、TP-73水平变化可反映替雷利珠单抗联合TP化疗方案在肺癌治疗中的效果,在综合疗效评估中有较高的应用价值。

低剂量CT结合SHOX2、RASSF1A甲基化在肺癌早期预警中的应用

目的探讨低剂量CT结合Ras相关区域家族蛋白1A(RASSF1A)、矮小同源盒基因2(SHOX2)甲基化在肺癌早期预测中的应用价值。方法选取2021年1月~2023年1月我院90例拟行肺结节手术患者,根据手术病理学分为肺良性结节组和肺癌组。2组均于术前行低剂量CT检查、SHOX2、RASSF1A甲基化检测,采用Kappa指数分析上述检查结果与手术病理学一致性,分析低剂量CT、SHOX2、RASSF1A甲基化与血清肿瘤标志物[癌胚抗原(CEA)、神经元特异性烯醇化酶(NSE)、鳞状细胞癌抗原(SCC-Ag)、细胞角蛋白19片段(CYFRA21)]对肺癌诊断效能,采用Spearman低剂量CT检查、SHOX2、RASSF1A甲基化与临床病理特征相关性。结果低剂量CT、SHOX2、RASSF1甲基化及三者联合分别确定40例、43例、46例、58例肺癌,三者联合与手术病理学诊断肺癌效能一致性Kappa值为0.951;三者联合诊断肺癌敏感度96.67%、准确度97.78%均高于三者单一诊断效能(P<0.05);肺癌患者血清CEA、SCC、NSE、CYFRA21水平均高于肺良性结节患者(P<0.05);低剂量CT联合SHOX2、RASSF1甲基化诊断肺癌效能的AUC为0.983,近似于四种血清肿瘤标志物诊断肺癌效能的AUC 0.933;不同肿瘤直径、临床分期、组织学分化肺癌患者低剂量CT检出率及SHOX2、RASSF1A甲基化阳性率比较差异有统计学意义(P<0.05);肺癌患者低剂量CT检出率、SHOX2及RASSF1A甲基化阳性率与肿瘤直径、临床分期呈正相关,与组织学分化呈负相关(P<0.05)。结论低剂量CT联合SHOX2及RASSF1A甲基化可用于肺癌早期预警中,临床可通过其进行早期诊断、评估病情进展程度,以针对性展开后续治疗,改善预后。

EGFR突变NSCLC组织LncRNA TCF7L2表达及临床病理特征和预后分析

目的 探究长链非编码RNA(LncRNA)转录因子7类似物2(TCF7L2)在表皮生长因子受体基因(EGFR)突变的非小细胞肺癌(NSCLC)组织表达及其与病人临床病理特征和预后相关性。方法 选取2019年3月—2021年6月河北北方学院附属第一医院治疗的EGFR突变NSCLC病人104例为研究对象,分别取其癌组织和癌旁组织应用逆转录PCR(RT-PCR)检测LncRNA TCF7L2的表达,比较不同组织TCF7L2表达及其与临床病理特征和预后的相关性。结果 RT-PCR检测结果显示,癌组织中LncRNA TCF7L2表达量显著高于癌旁组织(t=12.410,P<0.05)。与LncRNA TCF7L2低表达病人比较,高表达者发生淋巴结转移更多、肿瘤体积更大(χ^(2)=4.579、7.762,P<0.05);LncRNA TCF7L2高表达病人6、9和12个月的生存率较低,但差异均无统计学意义(P>0.05)。结论 LncRNA TCF7L2在EGFR突变NSCLC病人癌组织表达高于癌旁组织,且TCF7L2高表达病人的淋巴结转移风险较高、肿瘤体积较大,其生存率可能较低。

PD-1抑制剂联合一线化疗方案对晚期驱动基因阴性肺腺癌的效果及安全性分析

目的探讨程序性细胞死亡蛋白-1(PD-1)抑制剂联合一线化疗方案治疗晚期驱动基因阴性肺腺癌的效果及安全性。方法回顾性收集2019年1月至2021年12月收治的60例晚期驱动基因阴性的肺腺癌患者病历资料,依据治疗方式不同分组,将30例接受培美曲塞、顺铂联合PD-1抑制剂(信迪利单抗)治疗的病历资料纳入观察组另30例接受培美曲塞联合顺铂治疗的病历资料纳入对照组。对比2组患者治疗前、治疗30 d时2组临床疗效[客观缓解率(ORR)、疾病控制率(DCR)]、免疫功能[CD_(4)^(+)、CD_(8)^(+)、CD_(4)^(+)/CD_(8)^(+)]、肿瘤标志物[癌胚抗原(CEA)、细胞角蛋白19片段(CYFRA21-1)]、不良反应[恶心、粒细胞减少、肝功能异常、甲状腺功能异常、肺炎、皮疹、血小板降低]。结果观察组ORR显著高于对照组,差异有统计学意义(P<0.05);2组DCR比较差异无统计学意义(P>0.05);治疗后,观察组CD_(4)^(+)、CD_(4)^(+)/CD_(8)^(+)水平高于治疗前,且高于对照组(P<0.05);治疗后,2组CYFRA21-1、CEA水平均下降,且观察组低于对照组(P<0.05)。观察组甲状腺功能异常、肺炎、皮疹等发生例数显著多于对照组,差异有统计学意义(P<0.05)。结论PD-1抑制剂联合化疗一线治疗可有效改善晚期驱动基因阴性肺腺癌患者免疫功能,并降低肿瘤标志物水平,疗效确切。

GPR120基因通过调控NLRP3炎症小体激活保护脓毒症肺损伤的机制研究

目的通过脂多糖(Lipopolysaccharide,LPS)诱导的脓毒症模型验证G蛋白偶联受体120(G-protein coupled receptor120,GPR120)基因对NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)炎症小体及肺损伤的影响,并探索其调控分子机制。方法通过C57BL/6小鼠构建体内脓毒症模型,通过GPR120基因激动剂TUG891进行干预,验证GPR120基因对脓毒症小鼠肺损伤的保护作用;然后进行转录组测序,筛选差异信号通路,并在动物模型中验证NLRP3炎症小体及调控蛋白的差异表达。通过慢病毒转染构建GPR120基因过表达/低表达的Raw264.7单核巨噬细胞株,观察GPR120基因对NLRP3炎症小体的调控作用。结果与脓毒症组相比,LPS+TUG891组小鼠肺组织中包括cAMP通路基因在内的77个基因表达显著上调,37个基因表达下降。LPS组的GPR120水平较正常对照组显著降低,同时cAMP/PKA信号通路关键蛋白CREB及PKA表达减少,NLRP3、Caspase-1及IL-1β等炎症小体激活相关蛋白水平升高(P<0.01),予以TUG891处理后,组织内GPR120表达回升,cAMP/PKA信号通路重新被激活(P<0.01),NLRP3炎症小体蛋白活化程度下降(P<0.05)。体外实验中,LPS诱导的脓毒症可引起细胞增殖活性下降,GPR120基因在脓毒症巨噬细胞中表达减低(P<0.001),通过干预GPR120基因表达,证实GPR120基因可负性调控NLRP3炎症小体的活化程度及细胞炎症反应(P<0.01)。结论脓毒症中GPR120基因的激活可通过抑制NLRP3炎症小体的活化,减轻脓毒症的炎症反应及肺损伤。

肺癌患者支气管肺泡灌洗液中DAPL1和MLH1基因甲基化水平检测及其临床意义研究

目的检测肺癌患者支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中凋亡相关蛋白激酶样蛋白1(death-associated protein kinase like 1,DAPL1)和错配修复基因(Mut L homologue l,MLH1)甲基化水平,进一步研究基因甲基化对早期肺癌的诊断价值及其与临床病理特征的关系。方法回顾性选取2022年1月~2024年1月蒙城县第二人民医院收治的疑似早期肺癌患者142例,根据最终病理学结果分为肺癌组(n=82)和肺部良性病变组(n=60)。采用实时定量聚合酶链反应(qRT-PCR)法检测两组BALF样本中DAPL1和MLH1甲基化水平;分析DAPL1和MLH1甲基化对早期肺癌的临床诊断价值及其与肺癌患者临床病理特征的关系。结果肺癌组患者BALF中DAPL1和MLH1基因甲基化水平分别为53.66%(44/82),56.10%(46/82),明显高于良性病变组的11.67%(7/60)和18.33%(11/60),差异具有统计学意义(χ^(2)=56.544,20.565,均P<0.05)。DAPL1和MLH1基因甲基化诊断早期肺癌的敏感度分别为53.66%(44/82)和56.10%(46/82),特异度分别为88.33%(53/60)和81.67%(49/60),准确度分别为68.31%(97/142)和66.90%(95/142);DAPL1甲基化联合MLH1甲基化诊断早期肺癌的敏感度和准确度分别为86.59%(71/82),85.92%(122/142),均高于单一指标(Z=24.411,16.450,均P<0.05)。肺癌患者BALF中DAPL1和MLH1基因甲基化水平与临床分期、吸烟史及淋巴结转移密切相关(χ^(2)=5.493,13.083;8.167,6.946;9.303,4.523,均P<0.05)。Spearman相关性显示,DAPL1和MLH1基因甲基化与肺癌患者临床分期、吸烟史及淋巴结转移均呈正相关(r=0.523,0.602;0.548,0.498;0.630,0.524,均P<0.05)。结论BALF中DAPL1和MLH1基因甲基化检测对于早期肺癌具有较高的临床诊断价值,且两者基因甲基化水平与肺癌患者病情进展及吸烟史有关。

急性肺损伤相关分子标志物的鉴定及临床意义探索

目的通过公共数据库筛选急性肺损伤(ALI)及急性呼吸窘迫综合征(ARDS)相关分子标志物,并探索其临床意义。方法利用基因表达综合数据库(GEO)中有关ALI/ARDS基因表达芯片研究的两个数据集GSE76293和GSE10474,通过STRING网站和Cytoscape软件对差异基因进行蛋白互作网络分析并筛选ALI/ARDS相关关键基因。采用A549细胞构建ALI模型,并通过转录组测序验证关键基因在细胞中的表达差异情况。结果2个GEO数据集中共筛选出共同上调基因27个,共同下调基因26个。主要参与抗原加工和外源抗原递呈、免疫受体活性调节、内质网膜构成等生物学功能,且与抗原加工、细胞分化等信号通路有关。蛋白互作网络分析共筛选出10个ALI/ARDS相关关键基因,分别为CD4、HLADQB1、CD74、HLA-DRA、FCGR2B、TOR1A、RELA、NME8、RNF19B、RHOB。细胞转录组测序结果显示,关键基因的上调或下调特征及表达差异情况与GEO数据集分析结果一致。结论CD4等关键基因可能参与ALI/ARDS发生、发展的生物学过程,是ALI/ARDS临床诊断及预后预测的潜在个体化分子标志物。

miR-451及靶基因在非小细胞肺癌中的表达与调控研究

目的探讨miR-451及靶基因PSMB8在非小细胞肺癌中的表达及其与相关细胞周期蛋白的相关性,为非小细胞肺癌的治疗提供新的靶点。方法选取2020年1月至2022年12月该院胸外科行肺癌根治术的6例肺癌患者组织标本,将其按癌组织和癌旁组织进行分组,各6份。比较2组miR-451、蛋白酶体β亚单位8(PSMB8)、细胞周期蛋白D1(Cyclin D1)、Cyclin E1、细胞周期素依赖激酶4(CDK4)、CDK2蛋白相对表达水平,分析miR-451与相关细胞周期蛋白的相关性。结果肺癌组织组miR-451的CT值高于癌旁组织组,差异有统计学意义(P<0.05)。肺癌组织组miR-451的2^(-ΔΔCT)值为0.53,癌旁组织组miR-451相对表达量是肺癌组织组的1.87倍。肺癌组织组PSMB8相对表达水平为(1.55±0.23),明显高于癌旁组织组的(0.98±0.15),差异有统计学意义(P<0.05)。肺癌组织组Cyclin D1、Cyclin E1、CDK4、CDK2蛋白相对表达水平明显高于癌旁组织组,差异有统计学意义(P<0.05)。miR-451相对表达水平与Cyclin D1、Cyclin E1、CDK4、CDK2蛋白相对表达水平呈负相关(r=-0.93、-0.89、-0.86、-0.84,P<0.05)。结论肺癌组织中miR-451低表达,可减少对靶基因PSMB8表达的抑制作用,进而干扰细胞周期相关蛋白的表达,影响细胞增殖和凋亡,从而间接调控肺癌的发生发展。

非小细胞肺癌组织LncRNA MIR503HG、Wnt1表达与患者术后5年内生存的相关性

目的 探究非小细胞肺癌(NSCLC)组织中长链非编码RNA(LncRNA)miR503宿主基因(MIR503HG)、Wnt1表达水平与患者术后5年内生存的关系。方法 纳入108例NSCLC患者,并于术中收集患者肺癌组织和癌旁组织。荧光定量PCR法检测肺癌及癌旁组织中LncRNA MIR503HG、Wnt1 mRNA表达水平;免疫组织化学染色检测肺癌及癌旁组织中Wnt1蛋白表达。对NSCLC患者术后随访5年,记录随访期内生存状况。比较癌旁组织、肺癌组织中LncRNA MIR503HG、Wnt1 mRNA和蛋白阳性表达情况;比较不同结局NSCLC患者肺癌组织中两者表达水平及其在不同临床病理特征中的差异;Pearson法分析肺癌组织中两者表达水平的相关性;Kaplan-Meier生存曲线分析肺癌组织中两者表达水平与患者术后5年内生存的关系;多因素Cox回归分析NSCLC患者术后5年内生存的影响因素;受试者工作特征(ROC)曲线评估肺癌组织LncRNA MIR503HG、Wnt1 mRNA表达水平对患者术后5年内生存的预测价值。结果 肺癌组织中LncRNA MIR503HG表达水平低于癌旁组织,Wnt1 mRNA表达水平和Wnt1蛋白阳性表达率高于癌旁组织(P<0.05)。低分化、TNM分期Ⅲ期、有淋巴结转移NSCLC患者肺癌组织LncRNA MIR503HG表达水平分别低于中高分化、TNM分期Ⅰ—Ⅱ期、无淋巴结转移NSCLC患者;Wnt1 mRNA表达水平分别高于中高分化、TNM分期Ⅰ—Ⅱ期、无淋巴结转移NSCLC患者(P<0.05)。肺癌组织中LncRNA MIR503HG与Wnt1 mRNA表达水平呈负相关(P<0.05)。108例NSCLC患者术后随访5年,生存48例(生存组),死亡60例(死亡组);LncRNA MIR503HG高表达组、Wnt1 mRNA低表达组术后5年累积生存率分别高于LncRNA MIR503HG低表达组和Wnt1 mRNA高表达组(P<0.05)。与生存组相比,死亡组肺癌组织LncRNA MIR503HG表达水平降低,Wnt1 mRNA表达水平升高(P<0.05)。肺癌组织LncRNA MIR503HG低表达、Wnt1 mRNA高表达、低分化、TNM分期为Ⅲ期、有淋巴结转移均是影响NSCLC患者术后5年内生存的独立危险因素(P<0.05)。肺癌组织LncRNA MIR503HG、Wnt1 mRNA及二者联合预测NSCLC患者术后5年内生存的曲线下面积(AUC)分别为0.823、0.728和0.885,联合预测效能更高(P<0.05)。结论NSCLC患者肺癌组织中LncRNA MIR503HG呈低表达,Wnt1呈高表达,二者与患者临床病理特征和术后5年内生存情况密切相关,对患者术后5年内生存情况有较高预测价值。

肺癌组织PTPN6、PAX8表达水平及其与患者预后的关系

目的:探讨蛋白质酪氨酸磷酸酶非受体型6(PTPN6)、配对盒基因8抗体(PAX8)在肺癌组织中表达水平及其与患者预后的关系。方法:选取本院2019年1月至2021年1月收治的86例肺癌患者,在术中收集癌组织和癌旁组织。采用实时荧光定量PCR(qRT-PCR)检测组织中PTPN6、PAX8的mRNA表达,采用免疫组化法检测组织中PTPN6、PAX8的蛋白表达。结果:PTPN6 mRNA在肺癌组织中的表达低于癌旁组织(P<0.05);PAX8 mRNA在肺癌组织中的表达高于癌旁组织(P<0.05);肺癌组织中PTPN6蛋白阳性表达的患者3年生存率高于阴性表达者(P<0.05);PAX8蛋白阳性表达的患者3年生存率低于阴性表达者(P<0.05);淋巴结转移、肿瘤直径>3.0 cm、低分化、TNM分期为Ⅲ~Ⅳ期、PTPN6阴性表达、PAX8阳性表达为影响肺癌患者预后的危险因素(P<0.05)。结论:肺癌组织PTPN6表达水平下降,PAX8表达水平上升,可作为评估患者预后的指标。

抗炎通腑方对脓毒症急性肺损伤大鼠mtDNA-STING-NLRP3信号通路的影响

目的探讨抗炎通腑方对脓毒症急性肺损伤(ALI)大鼠的影响及作用机制。方法将35只大鼠随机分为正常组、模型组、地塞米松(DEX)组、中药(TCM)组和TCM+DEX组,每组7只。分别给予相应药物灌胃、造模。酶联免疫吸附实验(ELISA)法检测白细胞介素(IL)-6、IL-1β、IL-18、肿瘤坏死因子-α(TNF-α)含量及血清中巨噬细胞计数;采用实时荧光定量多聚核苷酸链式反应(RT-qPCR)和蛋白免疫印迹(Western blot)检测肺组织中干扰素基因刺激因子(STING)、磷酸化干扰素基因刺激因子(P-STING)、NOD样受体热蛋白结构域相关蛋白3(NLRP3)、凋亡相关斑点样蛋白信使核糖核酸(ASC mRNA)和相关蛋白的表达水平。测定肺组织湿/干比(W/D),观察肺组织病理学改变。检测肺组织中巨噬细胞表达情况。结果与正常组比较,各组指标显著升高;与模型组比较,DEX组、TCM组及TCM+DEX组各指标均显著降低。与模型组比较,TCM+DEX组改变最为明显,肺组织损伤改善,W/D值(4.77±0.29 vs.3.78±0.48)及巨噬细胞计数(13.39±2.06 vs.7.09±1.42)均降低(P<0.05),IL-6(152.51±22.27 vs.58.92±13.53)、IL-1β(126.19±10.02 vs.45.69±6.67)、IL-18(59.12±6.31 vs.31.75±4.23)及TNF-α(126.57±8.25 vs.49.59±8.12)水平均降低(P均<0.01),肺组织中线粒体DNA(mtDNA)损伤及释放、P-STING(0.32±0.03 vs.0.16±0.03)、STING mRNA(19.24±2.70 vs.0.32±0.16)、NLRP3 mRNA(20.03±5.06 vs.1.20±0.04)、ASC mRNA(16.96±4.31 vs.3.41±2.52)和相关蛋白的表达均降低(P<0.01)。结论抗炎通腑方可能通过调控mtDNA-STING-NLRP3信号通路减轻脓毒症ALI大鼠的炎症。