目的分析沉默分化簇(CD)147对非小细胞肺癌(NSCLC)细胞生物学特征及上皮间质转化(EMT)的影响,并探究可能的相关分子机制。方法回顾性收集2023年8月~2024年3月空军军医大学唐都医院收治的30例NSCLC患者癌灶组织及癌旁正常组织样本,从中...目的分析沉默分化簇(CD)147对非小细胞肺癌(NSCLC)细胞生物学特征及上皮间质转化(EMT)的影响,并探究可能的相关分子机制。方法回顾性收集2023年8月~2024年3月空军军医大学唐都医院收治的30例NSCLC患者癌灶组织及癌旁正常组织样本,从中科院细胞库购买人正常肺上皮细胞BEAS-2B及NSCLC细胞系A549、NCI-H460、NCI-H1299,采用实时荧光定量PCR(qRT-PCR)法检测NSCLC癌灶组织和癌旁正常组织,以及各细胞系中CD147 mRNA表达水平。将NCI-H460细胞分为control组、si-NC组、si-CD147组、SC-79组及si-CD147+SC-79组,通过CCK-8法、Transwell法及流式细胞仪检测敲低CD147对NSCLC细胞增殖、迁移、凋亡的影响。蛋白免疫印迹(Western blot)检测EMT相关标志物神经钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、上皮钙黏附蛋白(E-cadherin)及锌指转录因子(Slug)、扭曲蛋白(Twist)表达,以及磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)通路相关蛋白表达。结果NSCLC组织中CD147 mRNA(1.64±0.25)表达水平明显高于癌旁正常组织(0.43±0.06),差异具有统计学意义(t=8.152,P<0.05)。NSCLC细胞中CD147 mRNA表达水平较人正常肺上皮细胞明显升高(1.24±0.13、3.31±0.37、2.16±0.29 vs 0.35±0.04),差异具有统计学意义(F=80.516,P<0.05)。与control组相比,si-CD147组细胞增殖率(53.74%±4.07%vs 78.92%±6.23%)、迁移率(19.57%±2.69%vs 37.24%±3.12%)明显降低,细胞凋亡率(39.04%±2.74%vs 16.14%±2.14%)明显升高(t=5.548、7.068、12.036),N-cadherin(0.41±0.02 vs 1.01±0.02)、Vimentin(0.36±0.04 vs 1.02±0.04)蛋白表达降低,E-cadherin(1.85±0.14 vs 0.98±0.02)蛋白表达升高,Slug(0.49±0.03 vs 1.02±0.03)和Twist(0.53±0.05 vs 1.03±0.04)蛋白水平明显抑制(t=12.766~36.742),p-PI3K(0.42±0.03 vs 1.01±0.03)、p-AKT(0.45±0.03 vs 1.01±0.02)和p-mTOR(0.38±0.02 vs 0.99±0.02)蛋白表达水平明显降低(t=8.377、7.286、9.694),差异具有统计学意义(均P<0.05)。与SC-79组相比,si-CD147+SC-79组p-PI3K、p-AKT、p-mTOR蛋白表达明显升高(t=11.075、14.573、13.666),促进细胞增殖及迁移、降低细胞凋亡、促进EMT过程(t=2.834~11.396),差异具有统计学意义(均P<0.05);SC-79可逆转敲低CD147对PI3K/AKT/mTOR通路的影响(t=11.643、8.848、10.965)和PI3K/AKT/mTOR通路相关蛋白的表达(t=2.595~14.476),差异具有统计学意义(均P<0.05)。结论CD147高表达可通过PI3K/AKT/mTOR通路诱导EMT过程,促进肿瘤细胞的增殖及迁移,抑制细胞凋亡,参与NSCLC的发生发展。展开更多
Pediatric cancers are particularly significant due to their uncommon occurrence in children,driven by a variety of underlying factors.Because of their distinct molecular and genetic makeup,which makes early detection ...Pediatric cancers are particularly significant due to their uncommon occurrence in children,driven by a variety of underlying factors.Because of their distinct molecular and genetic makeup,which makes early detection challenging,they are linked to problems.Diagnostic methods like imaging and tissue biopsy are only effective when the tumor has reached a size that can be identified.The liquid biopsy technique,the least intrusive and most convenient diagnostic method,is the subject of this review.It focuses on the significance of single cell analysis in examining uncommon cancer types.The many biomarkers found in bodily fluids and the cancer types they are linked to in children have been assessed,as has the potential route towards early detection and cancer recurrence forecasting.Combining the single cell liquid biopsy with the newest technologies,such as computational and multi-omics approaches,which have improved the efficiency of processing massive and unique genetic data,appears promising.This article discusses on a number of case reports for uncommon pediatric malignancies,such as Neuroblastoma,Medulloblastoma,Wilms Tumor,Rhabdomyosarcoma,Ewing Sarcoma,and Retinoblastoma,as well as their liquid biopsy profiles.Furthermore,the findings raise ethical questions regarding the therapeutic application of the technology as well as possible difficulties related to clinical translation.The likelihood that this single cell liquid biopsy will be clinically validated and eventually used as a routine diagnostic tool for uncommon pediatric cancers will rise with the realistic approach to sensitivity monitoring,specificity upgrading,and optimization.展开更多
目的分析甲基转移酶3(METTL3)抑制剂STM2457对人肝癌细胞系HepG2的影响,重点研究其对N6-甲基腺苷(m6A)表达的影响及其抗肿瘤机制。方法将HepG2细胞分为实验组(STM2457处理)和对照组(DMSO处理)。利用纳米孔(Nanopore)测序技术,结合m6Anet...目的分析甲基转移酶3(METTL3)抑制剂STM2457对人肝癌细胞系HepG2的影响,重点研究其对N6-甲基腺苷(m6A)表达的影响及其抗肿瘤机制。方法将HepG2细胞分为实验组(STM2457处理)和对照组(DMSO处理)。利用纳米孔(Nanopore)测序技术,结合m6Anet,NanoCount,xPore和GFOLD方法,分别对m6A修饰水平、转录组表达及差异基因进行分析。通过基因本体(GO)和京都基因与基因组百科(KEGG)对差异基因进行功能富集分析。结果STM2457降低HepG2细胞的m6A修饰位点数量(6446 vs 11549)及修饰水平(0.95±0.03 vs 0.98±0.03),差异具有统计学意义(Z=-19.915,P<0.01)。差异基因分析共筛选出109个上调基因和340个下调基因,其中与肝癌发生发展密切相关的基因PDLIM5、AZGP1和RNASET2,其m6A修饰水平降低,而基因表达水平升高。功能富集分析结果显示,差异基因主要富集在细胞黏附、凋亡、翻译调控及肝细胞癌相关通路。结论STM2457通过抑制METTL3活性,降低HepG2细胞的m6A修饰水平,上调基因PDLIM5,AZGP1和RNASET2的表达,促进HepG2细胞凋亡,为肝癌治疗提供潜在治疗靶点。展开更多
BACKGROUND Early screening,preoperative staging,and diagnosis of lymph node metastasis are crucial for improving the prognosis of gastric cancer(GC).AIM To evaluate the diagnostic value of combined multidetector compu...BACKGROUND Early screening,preoperative staging,and diagnosis of lymph node metastasis are crucial for improving the prognosis of gastric cancer(GC).AIM To evaluate the diagnostic value of combined multidetector computed tomography(MDCT)and gastrointestinal endoscopy for GC screening,preoperative staging,and lymph node metastasis detection,thereby providing a reference for clinical diagnosis and treatment.METHODS In this retrospective study clinical and imaging data of 134 patients with suspected GC who were admitted between January 2023 and October 2024 were initially reviewed.According to the inclusion and exclusion criteria,102 patients were finally enrolled in the analysis.All enrolled patients had undergone both MDCT and gastrointestinal endoscopy examinations prior to surgical intervention.Preoperative clinical staging and lymph node metastasis findings were compared with pathological results.RESULTS The combined use of MDCT and gastrointestinal endoscopy demonstrated a sensitivity of 98.53%,specificity of 97.06%,accuracy of 98.04%,positive predictive value of 98.53%,and negative predictive value of 97.06%for diagnosing GC.These factors were all significantly higher than those of MDCT or endoscopy alone(P<0.05).The accuracy rates of the combined approach for detecting clinical T and N stages were 97.06%and 92.65%,respectively,outperforming MDCT alone(86.76% and 79.41%)and endoscopy alone(85.29% and 70.59%)(P<0.05).Among 68 patients with confirmed GC,50(73.53%)were pathologically diagnosed with lymph node metastasis.The accuracy for detecting lymph node metastasis was 66.00%with endoscopy,76.00%with MDCT,and 92.00% with the combined approach,all with statistically significant differences(P<0.05).CONCLUSION The combined application of MDCT and gastrointestinal endoscopy enhanced diagnostic accuracy for GC,provided greater consistency in preoperative staging,and improved the detection of lymph node metastasis,thereby demonstrating significant clinical utility.展开更多
文摘目的分析沉默分化簇(CD)147对非小细胞肺癌(NSCLC)细胞生物学特征及上皮间质转化(EMT)的影响,并探究可能的相关分子机制。方法回顾性收集2023年8月~2024年3月空军军医大学唐都医院收治的30例NSCLC患者癌灶组织及癌旁正常组织样本,从中科院细胞库购买人正常肺上皮细胞BEAS-2B及NSCLC细胞系A549、NCI-H460、NCI-H1299,采用实时荧光定量PCR(qRT-PCR)法检测NSCLC癌灶组织和癌旁正常组织,以及各细胞系中CD147 mRNA表达水平。将NCI-H460细胞分为control组、si-NC组、si-CD147组、SC-79组及si-CD147+SC-79组,通过CCK-8法、Transwell法及流式细胞仪检测敲低CD147对NSCLC细胞增殖、迁移、凋亡的影响。蛋白免疫印迹(Western blot)检测EMT相关标志物神经钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、上皮钙黏附蛋白(E-cadherin)及锌指转录因子(Slug)、扭曲蛋白(Twist)表达,以及磷脂酰肌醇-3-激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素靶蛋白(mTOR)通路相关蛋白表达。结果NSCLC组织中CD147 mRNA(1.64±0.25)表达水平明显高于癌旁正常组织(0.43±0.06),差异具有统计学意义(t=8.152,P<0.05)。NSCLC细胞中CD147 mRNA表达水平较人正常肺上皮细胞明显升高(1.24±0.13、3.31±0.37、2.16±0.29 vs 0.35±0.04),差异具有统计学意义(F=80.516,P<0.05)。与control组相比,si-CD147组细胞增殖率(53.74%±4.07%vs 78.92%±6.23%)、迁移率(19.57%±2.69%vs 37.24%±3.12%)明显降低,细胞凋亡率(39.04%±2.74%vs 16.14%±2.14%)明显升高(t=5.548、7.068、12.036),N-cadherin(0.41±0.02 vs 1.01±0.02)、Vimentin(0.36±0.04 vs 1.02±0.04)蛋白表达降低,E-cadherin(1.85±0.14 vs 0.98±0.02)蛋白表达升高,Slug(0.49±0.03 vs 1.02±0.03)和Twist(0.53±0.05 vs 1.03±0.04)蛋白水平明显抑制(t=12.766~36.742),p-PI3K(0.42±0.03 vs 1.01±0.03)、p-AKT(0.45±0.03 vs 1.01±0.02)和p-mTOR(0.38±0.02 vs 0.99±0.02)蛋白表达水平明显降低(t=8.377、7.286、9.694),差异具有统计学意义(均P<0.05)。与SC-79组相比,si-CD147+SC-79组p-PI3K、p-AKT、p-mTOR蛋白表达明显升高(t=11.075、14.573、13.666),促进细胞增殖及迁移、降低细胞凋亡、促进EMT过程(t=2.834~11.396),差异具有统计学意义(均P<0.05);SC-79可逆转敲低CD147对PI3K/AKT/mTOR通路的影响(t=11.643、8.848、10.965)和PI3K/AKT/mTOR通路相关蛋白的表达(t=2.595~14.476),差异具有统计学意义(均P<0.05)。结论CD147高表达可通过PI3K/AKT/mTOR通路诱导EMT过程,促进肿瘤细胞的增殖及迁移,抑制细胞凋亡,参与NSCLC的发生发展。
文摘Pediatric cancers are particularly significant due to their uncommon occurrence in children,driven by a variety of underlying factors.Because of their distinct molecular and genetic makeup,which makes early detection challenging,they are linked to problems.Diagnostic methods like imaging and tissue biopsy are only effective when the tumor has reached a size that can be identified.The liquid biopsy technique,the least intrusive and most convenient diagnostic method,is the subject of this review.It focuses on the significance of single cell analysis in examining uncommon cancer types.The many biomarkers found in bodily fluids and the cancer types they are linked to in children have been assessed,as has the potential route towards early detection and cancer recurrence forecasting.Combining the single cell liquid biopsy with the newest technologies,such as computational and multi-omics approaches,which have improved the efficiency of processing massive and unique genetic data,appears promising.This article discusses on a number of case reports for uncommon pediatric malignancies,such as Neuroblastoma,Medulloblastoma,Wilms Tumor,Rhabdomyosarcoma,Ewing Sarcoma,and Retinoblastoma,as well as their liquid biopsy profiles.Furthermore,the findings raise ethical questions regarding the therapeutic application of the technology as well as possible difficulties related to clinical translation.The likelihood that this single cell liquid biopsy will be clinically validated and eventually used as a routine diagnostic tool for uncommon pediatric cancers will rise with the realistic approach to sensitivity monitoring,specificity upgrading,and optimization.
文摘目的分析甲基转移酶3(METTL3)抑制剂STM2457对人肝癌细胞系HepG2的影响,重点研究其对N6-甲基腺苷(m6A)表达的影响及其抗肿瘤机制。方法将HepG2细胞分为实验组(STM2457处理)和对照组(DMSO处理)。利用纳米孔(Nanopore)测序技术,结合m6Anet,NanoCount,xPore和GFOLD方法,分别对m6A修饰水平、转录组表达及差异基因进行分析。通过基因本体(GO)和京都基因与基因组百科(KEGG)对差异基因进行功能富集分析。结果STM2457降低HepG2细胞的m6A修饰位点数量(6446 vs 11549)及修饰水平(0.95±0.03 vs 0.98±0.03),差异具有统计学意义(Z=-19.915,P<0.01)。差异基因分析共筛选出109个上调基因和340个下调基因,其中与肝癌发生发展密切相关的基因PDLIM5、AZGP1和RNASET2,其m6A修饰水平降低,而基因表达水平升高。功能富集分析结果显示,差异基因主要富集在细胞黏附、凋亡、翻译调控及肝细胞癌相关通路。结论STM2457通过抑制METTL3活性,降低HepG2细胞的m6A修饰水平,上调基因PDLIM5,AZGP1和RNASET2的表达,促进HepG2细胞凋亡,为肝癌治疗提供潜在治疗靶点。
文摘BACKGROUND Early screening,preoperative staging,and diagnosis of lymph node metastasis are crucial for improving the prognosis of gastric cancer(GC).AIM To evaluate the diagnostic value of combined multidetector computed tomography(MDCT)and gastrointestinal endoscopy for GC screening,preoperative staging,and lymph node metastasis detection,thereby providing a reference for clinical diagnosis and treatment.METHODS In this retrospective study clinical and imaging data of 134 patients with suspected GC who were admitted between January 2023 and October 2024 were initially reviewed.According to the inclusion and exclusion criteria,102 patients were finally enrolled in the analysis.All enrolled patients had undergone both MDCT and gastrointestinal endoscopy examinations prior to surgical intervention.Preoperative clinical staging and lymph node metastasis findings were compared with pathological results.RESULTS The combined use of MDCT and gastrointestinal endoscopy demonstrated a sensitivity of 98.53%,specificity of 97.06%,accuracy of 98.04%,positive predictive value of 98.53%,and negative predictive value of 97.06%for diagnosing GC.These factors were all significantly higher than those of MDCT or endoscopy alone(P<0.05).The accuracy rates of the combined approach for detecting clinical T and N stages were 97.06%and 92.65%,respectively,outperforming MDCT alone(86.76% and 79.41%)and endoscopy alone(85.29% and 70.59%)(P<0.05).Among 68 patients with confirmed GC,50(73.53%)were pathologically diagnosed with lymph node metastasis.The accuracy for detecting lymph node metastasis was 66.00%with endoscopy,76.00%with MDCT,and 92.00% with the combined approach,all with statistically significant differences(P<0.05).CONCLUSION The combined application of MDCT and gastrointestinal endoscopy enhanced diagnostic accuracy for GC,provided greater consistency in preoperative staging,and improved the detection of lymph node metastasis,thereby demonstrating significant clinical utility.
