THE EXPRESSIONS OF HBV X GENE AND ets-2, IGF-Ⅰ, c-myc AND N-ras ONCOGENES IN HUMAN HEPATOCELLULAR CARCINOMA AND TUMOR-ADJACENT TISSUES

The expressions of HBV X gene and ets-2, IGF-I, c-myc and N-ras were studied in 7 pairs of human primary hepatocellular carcinoma (PHC) and tumor-adjacent tissues, using RNA hybridization and im-munoblot methods. The results showed that specific 17 and 28 kD HBV X gene products (HBxAg) were existed in a portion of PHC and tumor-adjacent tissues. The 17 kD HBxAg was detected in the sera of 3 patients who also had 17 kD HBxAg in their liver tissues. Multiple expressions of oncogenes such as ets-2, c-myc and N-ras were observed in PHC and tumor-adjacent tissues that had HBxAg expressed, indicating HBxAg might function as a transactivator in the course of intracellular proto-oncogene activation. It is also observed that in some tumor-adjacnet tissues the expressions of ets-2, c-myc and N-ras were higher than those in corresponding PHC. The relationship of HBxAg to the expression of est-2, IGF-Ⅱ, c-myc and their possible roles in the carcinogenesis of PHC are discussed.

THE EXPRESSION OF C-MYC AND N-RAS ONCOGENES IN HUMAN HEPATOCELLULAR CARCINOMA-AN IN SITU HYBRIDIZATION STUDY ON PARAFFIN EMBEDDED TISSUE SECTIONS

In this research, we investigated the expression of C myc and N-ras mRNAs on 21 cases paraffin- embedded tissue sections of hepatocellular carcinoma(HCC) using insitu hybridization technique with biotinylated labelled cDNA probes. Of 21 cases of hepatoma , C-myc mRNA was positive-expressed in 9 cases(42. 9 % ) and N-ras positive in 4 cases ( 19% ) in hepatoma cells, and C-myc and N-ras positive in 4 and 1 cases respectively in peritumor hepatocytes. C- myc mRNAs were localized within cytoplasms of both hepatoma cells and peritumor hepatocytes. However , the positive intensities of C-myc and N-ros mRNAs in hepatoma cells were much greater than those in peritumor hepatocytes. The results indicated that Cmyc and N-ras oncogenes were overexpressed in HCC, and may play an important role in coordinatively maintaince of the malignant phenotypes in HCC.

乙肝相关肝癌患者血清中癌基因C-myc、N-ras、PLK1、FGF2蛋白水平对TACE术后预后的预测价值分析

目的探讨乙型肝炎(简称乙肝)相关肝癌患者血清中癌基因[增殖相关基因(C-myc)、转化基因(N-ras)、丝/苏氨酸激酶1(PLK1)、成纤维细胞生长因子2(FGF2)]蛋白水平对肝动脉化疗栓塞术(TACE)后预后的预测价值。方法选取该院于2016年7月至2021年1月收治的乙肝相关肝癌患者127例,根据随访结果将患者分为死亡组和存活组。采用双抗体夹心酶联免疫吸附法测定患者术前血清癌基因C-myc、N-ras、PLK1、FGF2蛋白水平,单因素及多因素Cox分析血清中癌基因C-myc、N-ras、PLK1、FGF2蛋白水平与乙肝相关肝癌患者TACE术后预后的危险因素;通过受试者工作特征曲线来评估血清癌基因C-myc、N-ras、PLK1、FGF2蛋白水平对乙肝相关肝癌患者预后的预测价值,并根据对应的截断值将乙肝相关肝癌患者分为血清中癌基因C-myc、N-ras、PLK1、FGF2蛋白水平高表达组与低表达组,采用Kaplan-Meier生存曲线评估不同血清癌基因C-myc、N-ras、PLK1、FGF2蛋白水平的预后生存情况。结果多因素Cox回归分析结果提示,TNM分期Ⅲ~Ⅳ期(HR=2.998,95%CI:1.239~7.257)、有门静脉转移(HR=3.737,95%CI:1.941~7.193)、有腹腔转移(HR=3.482,95%CI:1.709~7.097)、Child-Pugh分级B级(HR=2.587,95%CI:1.045~6.406)、高血清癌基因C-myc蛋白水平(HR=1.224,95%CI:1.090~1.374)、高血清癌基因N-ras蛋白水平(HR=1.218,95%CI:1.097~1.353)、高血清癌基因PLK1蛋白水平(HR=1.237,95%CI:1.110~1.379)、高血清癌基因FGF2蛋白水平(HR=1.141,95%CI:1.060~1.228)均是影响乙肝相关肝癌患者TACE术后预后的独立危险因素(均P<0.05)。血清癌基因C-myc、N-ras、PLK1、FGF2蛋白水平低表达组的总生存率均明显高于血清癌基因C-myc、N-ras、PLK1、FGF2蛋白水平高表达组,差异有统计学意义(均P<0.001)。结论血清癌基因C-myc、N-ras、PLK1、FGF2蛋白水平对乙肝相关肝癌患者TACE术后预后具有预测价值。

Tanshinone ⅡA improves Alzheimer’s disease via RNA nuclearenriched abundant transcript 1/microRNA-291a-3p/member RAS oncogene family Rab22a axis

BACKGROUND Alzheimer’s disease(AD)is a neurodegenerative condition characterized by oxidative stress and neuroinflammation.Tanshinone ⅡA(Tan-ⅡA),a bioactive compound isolated from Salvia miltiorrhiza plants,has shown potential neuroprotective effects;however,the mechanisms underlying such a function remain unclear.AIM To investigate potential Tan-ⅡA neuroprotective effects in AD and to elucidate their underlying mechanisms.METHODS Hematoxylin and eosin staining was utilized to analyze structural brain tissue morphology.To assess changes in oxidative stress and neuroinflammation,we performed enzyme-linked immunosorbent assay and western blotting.Additionally,the effect of Tan-ⅡA on AD cell models was evaluated in vitro using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay.Genetic changes related to the long non-coding RNA(lncRNA)nuclear-enriched abundant transcript 1(NEAT1)/microRNA(miRNA,miR)-291a-3p/member RAS oncogene family Rab22a axis were assessed through reverse transcription quantitative polymerase chain reaction.RESULTS In vivo,Tan-ⅡA treatment improved neuronal morphology and attenuated oxidative stress and neuroinflammation in the brain tissue of AD mice.In vitro experiments showed that Tan-ⅡA dose-dependently ameliorated the amyloid-beta 1-42-induced reduction of neural stem cell viability,apoptosis,oxidative stress,and neuroinflammation.In this process,the lncRNA NEAT1-a potential therapeutic target-is highly expressed in AD mice and downregulated via Tan-ⅡA treatment.Mechanistically,NEAT1 promotes the transcription and translation of Rab22a via miR-291a-3p,which activates nuclear factor kappa-B(NF-κB)signaling,leading to activation of the pro-apoptotic B-cell lymphoma 2-associated X protein and inhibition of the anti-apoptotic B-cell lymphoma 2 protein,which exacerbates AD.Tan-ⅡA intervention effectively blocked this process by inhibiting the NEAT1/miR-291a-3p/Rab22a axis and NF-κB signaling.CONCLUSION This study demonstrates that Tan-ⅡA exerts neuroprotective effects in AD by modulating the NEAT1/miR-291a-3p/Rab22a/NF-κB signaling pathway,serving as a foundation for the development of innovative approaches for AD therapy.

Concomitant epidermal growth factor receptor mutation/c-ros oncogene 1 rearrangement in non-small cell lung cancer: A case report

BACKGROUND Epidermal growth factor receptor(EGFR)mutation and c-ros oncogene 1(ROS1)rearrangement are key genetic alterations and predictive tumor markers for non-small cell lung cancer(NSCLC)and are typically considered to be mutually exc-lusive.EGFR/ROS1 co-mutation is a rare event,and the standard treatment appr-oach for such cases is still equivocal.CASE SUMMARY Herein,we report the case of a 64-year-old woman diagnosed with lung adenocar-cinoma,with concomitant EGFR L858R mutation and ROS1 rearrangement.The patient received two cycles of chemotherapy after surgery,but the disease prog-ressed.Following 1-month treatment with gefitinib,the disease progressed again.However,after switching to crizotinib,the lesion became stable.Currently,crizotinib has been administered for over 53 months with a remarkable treatment effect.CONCLUSION The efficacy of EGFR tyrosine kinase inhibitors and crizotinib was vastly different in this NSCLC patient with EGFR/ROS1 co-mutation.This report will aid future treatment of such patients.

1889例结直肠癌MSI、K-ras、N-ras和B-raf基因状态以及临床病理特征分析

目的通过对1889例结直肠癌微卫星不稳定(MSI)、K-ras、N-ras和B-raf基因检测,分析其基因状态与临床病理特征之间的关联性。方法收集东部战区总医院病理科结直肠癌患者手术切除大标本共计1889例,通过PCR荧光法联合毛细管电泳法对1889例结直肠癌进行MSI、K-ras、N-ras和B-raf检测,分析其基因状态与临床病理特征之间的关系,以及MSI与K-ras、N-ras和B-raf之间的关联性分析,并且对MSI自身位点的相关性分析。结果1889例结直肠癌患者中,微卫星高度不稳定(MSI-H)更易发生于右侧结肠(16.4%)、黏液腺癌(14.9%)、分化程度较低(18.7%)、年龄偏低(8.3%)、淋巴结未转移(9.1%)以及B-raf突变(5.2%)的患者中,K-ras突变更易发生于右侧结肠(50.7%)、黏液腺癌(50.7%)、分化程度较高(52.1%)、微卫星低度不稳定或稳定型(42.2%)的患者中,而B-raf突变更易发生在MSI-H(6.9%)的患者中,而N-ras基因则与临床病特征关联性不强。结论结直肠癌患者的MSI、K-ras和B-raf基因状态与临床病理特征存在着高度的关联性,因此通过分析其基因状态与临床病理特征之间的关系可以为临床靶向治疗以及预后提供更加可靠的依据。

TCGA-based analysis of oncogenic signaling pathways underlying oral squamous cell carcinoma

Background:Oral squamous cell carcinoma(OSCC)represents a prevalent malignancy in the oral and maxillofacial area,having a considerable negative impact on both the quality of life and overall survival of affected individuals.Our research endeavors to leverage bioinformatic approaches to elucidate oncogenic signaling pathways,with the ultimate goal of gaining deeper insights into the molecular underpinnings of OSCC pathogenesis,and thus laying the groundwork for the development of more effective therapeutic and preventive strategies.Methods:Differential expression analysis was performed on mRNA data from tumor and normal tissue groups to identify genes associated with OSCC,using The Cancer Genome Atlas database.Predictions of oncogenic signaling pathways linked to differentially expressedmRNAs were made,and these results were presented visually using R software,using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichments.Results:GO and KEGG analyses of 2938 differentially expressed genes in OSCC highlighted their significant involvement in various biological processes.Notably,these processes were related to the extracellular matrix,structural organization,connective tissue development,and cell cycle regulation.Conclusions:The comprehensive exploration of gene expression patterns provides valuable insights into potential oncogenic mechanisms in OSCC.

甲胎蛋白对HeLa细胞N-ras、p53和p21^(ras)表达的促进作用

大量研究已证明甲胎蛋白 (alpha fetoprotein ,AFP)对肿瘤细胞的增殖具有调节作用 .为探讨AFP对细胞生长促进作用的分子机理 ,采用从人脐带血中提取的AFP作用于体外培养的HeLa细胞 ,用Northern印迹分析法分析不同作用时间时细胞N rasmRNA的表达以及用Western印迹分析法分析p5 3、p2 1ras的表达 .结果发现 ,在AFP(2 0mg L)作用后 ,HeLa细胞的N rasmRNA、p5 3蛋白质和p2 1ras蛋白质的表达量与对照组比较在 12h和 2 4h时都有明显增加 .AFP的作用均可被抗AFP单克隆抗体所拮抗 .实验结果提示 ,AFP对细胞生长的调节作用可能通过促进这些原癌基因的表达来实现 .

含人N-ras反意基因的假型逆转录病毒对裸小鼠体内人肝癌移植瘤生长和N-ras表达的作用

含人反意基因的假型逆转录病毒已在许多实验室构建,并用于抑制病毒的或细胞的基因表达。抑制基因活性是一个新的遗传分析方法,其机制是通过一条互补于正常RNA的反意链与有意义链的结合而影响其功能,它的优点是能根据目的基因的转录产物设制有效的反意RNA,研究该基因在细胞中的生物学作用。RNA逆转录病毒因其独特性结构和繁衍形式,已成为理想的基因转移载体。

树舌多糖GF对HepA瘤细胞N-ras、C-myc基因表达的影响

目的为探明树舌多糖GF对HepA瘤组织N-ras、C-myc基因表达的影响。方法运用原位杂交法、SP免疫组化染色技术及多媒体彩色病理图文分析系统测定HepA瘤细胞中N-ras、C-myc mRNA及蛋白的表达量。结果树舌多糖组、猪苓多糖组中N-ras、C-myc mRNA及蛋白的表达量均显著低于荷瘤组(P<0.01),树舌多糖组与猪苓多糖组无显著性差异。结论树舌多糖GF可通过降低N-ras、C-myc mRNA及蛋白的表达,而抑制HepA瘤细胞的增殖。

益气养阴方及其拆方影响急性髓细胞白血病细胞Flt3和N-ras表达研究

目的研究益气养阴方及其拆方后的扶正方、祛邪方对Flt3和N-ras在人急性髓细胞白血病(acute myeloid leukemia,AML)表达的影响,探讨益气养阴方治疗白血病的作用机制。方法收集60例AML患者骨髓单个核细胞,将所提取每例患者的细胞混悬液分成4组:对照组不加药物,实验组分别加入益气养阴方、扶正方、祛邪方中药制剂。采用逆转录-聚合酶链反应(RT-PCR),免疫印迹法(Western bloting)观察益气养阴组、扶正组、祛邪组及对照组对Flt3、N-ras基因及FLT3蛋白表达的影响。结果 RT-PCR法检测:对照组、益气养阴组、扶正组、祛邪组Flt3基因表达率分别为(90.78±6.92)%、(38.18±4.50)%、(65.57±5.55)%、(61.35±6.39)%,N-ras基因表达率分别为(93.28±5.54)%、(34.38±6.69)%、(59.42±7.35)%、(65.28±7.64)%,益气养阴组、扶正组、祛邪组与对照组比较,差异均有统计学意义(P<0.05)。Westernbloting检测:对照组、益气养阴组、扶正组、祛邪组FLT3蛋白灰度值分别为0.8127±0.0284、0.4265±0.0353、0.5396±0.0274、0.5473±0.0282,对照组与益气养阴组、扶正组、祛邪组比较差异有统计学意义(P<0.01)。结论中药益气养阴方可抑制AML细胞的克隆性增殖,可降低Flt3和N-ras在AML细胞的表达水平,对AML治疗具有作用。

IGF-Ⅱ对肝癌细胞MHCC97-H癌基因C-myc和N-ras表达的影响

目的观察胰岛素样生长因子Ⅱ(IGF-Ⅱ)对肝癌细胞MHCC97-H中癌基因C-myc和N-ras表达的影响。方法培养肝癌细胞株MHCC97-H,用IGF-Ⅱ(50ng/mL)作用48h后,分别采用细胞免疫荧光、Western blot法及Real-time PCR法检测细胞中C-myc和N-ras的表达情况,并与对照组比较,进行定量分析。结果 C-myc及N-ras蛋白阳性表达主要位于细胞核。对照组、IGF-Ⅱ组MHCC97-H细胞中C-myc荧光表达量分别为(100.00±2.89)%、(254.00±35.57)%,N-ras荧光表达量分别为(100.00±14.43)%、(257.30±22.43)%。与对照组相比,IGF-Ⅱ组MHCC97-H细胞中C-myc及N-ras蛋白表达明显增强,差异有统计学意义(P<0.05)。与对照组相比,IGF-Ⅱ组MHCC97-H细胞中C-myc和N-ras的mRNA表达明显增加,差异有统计学意义(P<0.05)。结论 IGF-Ⅱ可上调肝癌细胞MHCC97-H中癌基因C-myc和N-ras的蛋白及mRNA表达,这可能是IGF-Ⅱ促进肝癌细胞增殖的分子机制之一,这为临床上肝癌防治提供了新线索。

中药复方对急性髓细胞性白血病干细胞Flt3和N-ras基因表达的影响(英文)

背景:在白血病患者体内存在一群白血病干细胞,这些恶性干细胞是白血病细胞存在和不断增殖的根源,针对干预白血病干细胞的特异性靶向治疗药物已成为近年来研究热点。目的:探讨中药复方对急性髓细胞性白血病干细胞Flt3和N-ras基因表达的影响。设计、时间及地点:随机区组实验,于2007-09/2008-12在山东中医药大学附属医院完成。对象:2006-2007年山东中医药大学附属医院、山东大学齐鲁医院血液科就诊的急性髓细胞性白血病初治患者50例,FAB分型M15例,M215例,M49例,M521例。方法:原方:黄芪、白花蛇舌草、小蓟、太子参、半枝莲、蒲公英、生地、黄精、女贞子、旱莲草、天冬、麦冬、白术、茯苓、甘草。扶正方:黄芪、太子参、生地、黄精、女贞子、天冬、麦冬、白术、茯苓、甘草。祛邪方:白花蛇舌草、小蓟、半枝莲、蒲公英、旱莲草。制剂:上述3组组方药物由山东中医药大学附属医院中药制剂实验室(国家三级重要制剂实验室)配制成1g/mL的药液,无菌试验阴性后过滤除菌备用。常规无菌采集急性髓细胞性白血病M1,M2,M4,M5型患者骨髓各5mL,稀释后加入淋巴细胞分离液,联合应用免疫磁珠分选系统和流式细胞仪分离纯化白血病干细胞。调整细胞浓度至2×108L-1,均分成4组:对照组不加药物,实验组分别加入原方、扶正方、祛邪方的对应中药制剂,终浓度为100mg/L,继续培养48h后进行指标检测。主要观察指标:RT-PCR法检测Flt3、N-ras基因的表达。结果:与对照组比较,原方组、扶正方组、祛邪方组Flt3表达阳性率均明显降低(P<0.05),但原方组降低幅度明显大于扶正方组、祛邪方组(P<0.05);扶正方组与祛邪方组Flt3表达阳性率比较无明显差异(P>0.05)。4组N-ras表达阳性率与Flt3表达情况基本一致。结论:Flt3和N-ras基因在白血病干细胞中存在过度表达,中药原方及其拆方后的扶正方、祛邪方均能够降低白血病干细胞中Flt3和N-ras基因的表达水平。

PCR-SSCP检测急性白血病患者N-ras基因突变及临床意义初探

目的 探讨N ras基因突变与急性白血病发生的关系。方法 采取骨髓和外周血白细胞提取DNA ,用PCR SSCP分析技术分别对急性白血病患者、正常对照组的N ras基因突变进行检测。结果 84例急性白血病患者中 2 5例发生N ras基因突变 ,基因变异频率为 2 9 8% ,其中急性淋巴细胞白血病为 18 6% ,急性髓细胞白血病为 41 4% ;11例随访 6个月 ,7例N ras基因突变消失 ,4例N ras基因突变持续存在 ;正常对照组未发现N ras基因突变。结论 N

DDPH 对实验性心肌肥厚大鼠左心室 N-ras mRNA 及蛋白质表达的影响

目的：研究１－（２，６－二甲基苯氧基）－２－（３，４－二甲氧基苯乙氨基）－丙烷盐酸盐（ＤＤＰＨ）对心肌肥厚大鼠左室Ｎ－ｒａｓｍＲＮＡ及蛋白质表达的影响。方法：用部分狭窄腹主动脉方法造成大鼠心肌肥厚模型，从术后第４周开始，ｉｇ不同剂量的ＤＤＰＨ（２５和５０ｍｇ·ｋｇ－１·ｄ－１），持续８ｗｋ。用ＲＮＡ狭缝杂交及免疫组化的方法检测Ｎ－ｒａｓｍＲ－ＮＡ及蛋白质表达的变化。结果：术后１２ｗｋ，发现肥厚组心肌组织Ｎ－ｒａｓｍＲＮＡ表达是对照组的３．１倍，蛋白质表达是对照组的２．９倍，而二个给药组ｍＲＮＡ及蛋白质的表达明显低于肥厚组，但仍高于对照组。结论：ＤＤＰＨ对大鼠腹主动脉部分狭窄所致心肌肥厚时Ｎ－ｒａｓｍＲＮＡ及蛋白质表达的增加有一定的逆转作用。

肝癌中医证型与野生型p53mRNA、N-ras蛋白表达相关性研究

目的:观察肝癌中医证型与野生型p53mRNA、N-ras表达相关性。方法:选择肝癌脾虚证36例和湿热证患者24例,应用原位杂交和免疫组化方法分别检测其肝癌组织野生型p53mRNA和N-ras蛋白表达,比较2组之间的差异。结果:脾虚证组野生型p53mRNA表达水平显著低于湿热证组(P<0．05);脾虚证组N-ras蛋白表达水平略低于湿热证组(P>0．05)。结论:野生型p53mRNA表达与旰癌中医证型有关,野生型p53 mRNA低表达是区别脾虚证和湿热证的特征之一。

骨肉瘤N-ras、c-myc基因异常及其蛋白产物的表达

目的：探讨癌基因Ｎ－ｒａｓ、ｃ－ｍｙｃ及其蛋白表达与骨肉瘤的关系。方法：Ｓｏｕｔｈｅｒｎｂｌｏｔ和免疫组织化学（ＬＳＡＢ法）同步检测９例人骨肉瘤Ｎ－ｒａｓ、ｃ－ｍｙｃ癌基因和其ｐ２１ｒａｓ、ｃ－ｍｙｃ蛋白产物表达。结果：Ｎ－ｒａｓ基因有１例为缺失改变，检出率１１％（１／９），ｃ－ｍｙｃ基因有３例为扩增改变，检出率３３％（３／９）。ｐ２１ｒａｓ、ｃ－ｍｙｃ基因蛋白表达阳性率分别为７８％（７例）、８９％（８例）。ｃ－ｍｙｃ基因蛋白在骨肉瘤的表达强度为最高。在９例骨肉瘤中，两种原癌基因蛋白共同表达６例，占７０％。结论：癌基因蛋白过度表达在骨肉瘤是一个常见现象，并从癌基因和其蛋白水平上提示癌基因Ｎ－ｒａｓ、ｃ－ｍｙｃ和ｐ２１ｒａｓ。

滋养细胞肿瘤N-ras基因原位分子杂交研究

目的 :为探讨滋养细胞肿瘤发病原因。方法 :采用核酸原位分子杂交技术 ,用地高辛标记的N ras基因探针与 40例滋养细胞肿瘤患者的刮宫标本及子宫切除标本进行原位杂交。用图像分析仪分别测各组阳性信号的灰度值作定量分析。结果 :随着恶性程度的增加 ,N ras基因阳性信号的表达呈增高趋势 ,恶性葡萄胎和绒毛膜癌明显高于良性葡萄胎和正常绒毛 ,有极显著性差异 (P <0 .0 1 )。结论 :N ras基因与滋养细胞的增殖、分化、恶变有关 ,由于N ras基因的过度表达而致恶变。

多发性骨髓瘤患者中N-ras基因突变的检测及其意义

目的 了解多发性骨髓瘤 (multiplemyeloma ,MM)患者中N ras基因突变率及临床意义。方法 采用聚合酶链反应 单链构象多态性银染法检测MM患者的N ras基因突变 ,并结合临床分析。结果 14例MM患者中 ,4例在N ras第 6 1密码子 ,1例在第 12、13密码子处发生突变 ,突变率为 35 .7%。突变者中 4例已死亡 ,其中 2例检测突变时已处于疾病终末期。结论 N ras基因在MM中有较高的突变率 ,对其的检测有助于病程进展和预后的分析。