Preliminary Study on c-Ha-ras Oncogene Mutations in Hydatidiform Mole Tissues

ve To study the presence of c-Ha-ras oncogene mutations in hydatidiform mole (HM) tissues and to further explore its relationship with mole's malignancy

C-Ha-ras Oncogene in Oral Leukoplakia Tissues

The amplification and the G-T mutation at codon 12 of the C-Ha-ras oncogene in oral leukoplakia tissues were analyzed by using polymerase chain reaction (PCR) and molecular biologic technique. The results showed that these tissues had no amplification of Ha-ras oncogene. Only one case harbored G-T mutation in 11 oral leukoplakias, but this mutation was absent in 10 normal oral mucosal tissues. The possible role and significance of C-Ha-ras oncogene in oral precancerous lesions were also discussed.

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.

The role of tazarotene-induced gene 1 in carcinogenesis:is it a tumor suppressor gene or an oncogene?

Tazarotene-induced gene 1(TIG1)is induced by a derivative of vitamin A and is known to regulate many important biological processes and control the development of cancer.TIG1 is widely expressed in various tissues;yet in many cancer tissues,it is not expressed because of the methylation of its promoter.Additionally,the expression of TIG1 in cancer cells inhibits their growth and invasion,suggesting that TIG1 acts as a tumor suppressor gene.However,in some cancers,poor prognosis is associated with TIG1 expression,indicating its protumor growth characteristics,especially in promoting the invasion of inflammatory breast cancer cells.This review comprehensively summarizes the roles of the TIG1 gene in cancer development and details the mechanisms through which TIG1 regulates cancer development,with the aim of understanding its various roles in cancer development.

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.

DIFFERENCES IN EXPRESSION AND REGULATION BETWEEN TRANSFORMED CELLS OF THE HUMAN GASTRIC CARCINOMA ONCOGENE Ha-ras AND THE UNTRANSFORMED PARENT CELLS

An Ha-ras oncogene was isolated from a cell line of gastric carcinoma called BGE-823 in order to elucidate genetic control and the influence of DNA sequences. The oncogene was cloned and identified as a single nucleotide substitution of thymine for guanine in the 12th codon through the sequencing of its first axon. We compared the differences of expression and regulation between the transformed Ha-ras cells and untransformed parent cells. Data indicated that the expression of Ha-ras in the transformed cells was five-fold higher than in the untransformed cells and that the Ha-ras gene in the former was hypersensitive toward DNase I. In addition, a nuclear protein of 35 kilodaltons bound strongly to the 2.5 Kb fragment located upstream of the 6.6 Kb Ha-ras gene and contained a CC rich region. These results suggest that there might be another mechanism of activation for the ras gene besides point mutation.

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.

中国人胃癌和正常组织基因组DNA中癌基因Ha-ras的限制性片段长度多态性研究

本文报道以PHS-49为探针,分析了中国人群中36例胃癌患者癌组织和25位正常人体组织基因组DNA中Ha-ras基因的BamHI限制性片段长度多态性(RFLPs)。发现了10种不同长度的片段和18种基因型。其中4种小于6kb的BamHI片段是迄今国外未见报道的,这可能是中国人群遗传多态性的一个特征。此外,在胃癌组织中发现Ha-ras的一些稀有等位基因和基因型的频率明显高于正常人群。对两名胃癌患者家系中的11名成员也作了RFLPs分析,发现有些成员出现3条和4条限制性片段的杂合个体,表明这些个体的染色体上含有的Ha-ras基因不只一份拷贝。对上述现象的可能原因作了分析和讨论。

鼻咽癌癌基因研究 Ⅱ.鼻咽癌癌基因属性-Ha-ras基因

用已知癌基因片段作为探针,检测经鼻咽癌CNE-2DNA二轮转染的NIH3T3转化细胞株,结果证实在转化细胞株中存在着与C-Ha-ras基因的同源基因,但该基因在带型和杂交强度上与CNE-2细胞DNA Ha-ras基因相比,没有大的改变。

蛋白激酶Cα对人正常肝和肝癌细胞中Ha-ras基因表达的影响

运用基因转染技术 ,将蛋白激酶C(PKCα)cDNA正向插入的真核表达重组质粒pXJ4 1 PKCα ,导入人正常肝细胞 (L 0 2 ) ,经蛋白质免疫印迹等检验 ,表明成功构建了稳定过表达PKCα的人正常肝细胞模型 (LT3) ,用LT3和表达反义PKCα的人肝癌细胞HT6为细胞模型 ,通过RT PCR ,蛋白质印迹 (Westernblot)等分析和进一步运用自行构建的真核表达质粒prasGL3,进行荧光素酶活性检测表明 :过表达PKCα可以促进L 0 2细胞的增殖速率 ,并引起Ha ras基因转录水平上升和Ha ras启动子活性升高 ;反之 ,表达反义PKCα的BEL 74 0 2细胞增殖被抑制 ,Ha ras基因转录水平下降 ,Ha ras启动子活性降低 .通过实验我们首次观察到 ,PKCα亚类对Ha ras癌基因表达的影响与其对Ha ras启动子活性的作用有关 ,PKCα可能参与了对Ha

PMA对人肝癌细胞中Ha-ras基因启动子活性的影响及与PKC活性的相关性

为探讨佛波脂PMA的生物作用与癌基因ras表达及与PKC信号通路的相关性,用人肝癌细胞BEL-7402为模型,以荧光素酶为报告基因观察了PMA对Ha-ras启动子活性的影响.当用100μg/L PMA长期处理人肝癌细胞BEL-7402时,可明显抑制肿瘤细胞生长,通过RT-PCR和Western Blot分析发现,PMA处理后的BEL-7402细胞中Ha-ras mRNA和蛋白水平均明显下降,进一步观察了PMA对Ha-ras基因启动子活性的影响。通过构建含有Ha-ras启动子序列的、以荧光素酶为报告基因的pGL3-ras真核表达载体,利用脂质体瞬时转染技术将其转入BEL-7402细胞中,并通过荧光素酶活性的检测,发现PMA作用48和72h后的BEL-7402细胞中,Ha-ras启动子活性分别被抑制了55％和54％,同时PKC活性分别下降了76.7％和75.7％。并进一步观察到PKCα和βⅠ蛋白水平明显降低。实验结果表明,PMA长期处理的人肝癌细胞可通过抑制Ha-ras启动子的活性使Ha-ras基因表达被抑制,并且可能与PKC信号通路有关。

实验性大鼠肝癌形成过程中c-myc、Ha-ras、Ki-ras的表达特点及其生物意义

目的：通过动态观察ｃ－ｍｙｃ、Ｈａ－ｒａｓ和Ｋｉ－ｒａｓ在实验性大鼠肝癌形成过程中的表达变化，探讨上述原癌基因与肝癌发生的关系及其生物学意义。方法：用３＇－Ｍｅ－ＤＡＢ饲喂ＳＤ大鼠，分别于第４，６，８，１４和１７周处死大鼠，取其肝脏，石蜡包埋，并进行核酸原位杂交。结果：在诱癌全过程中，ｃ－ｍｙｃ和Ｈａ－ｒａｓ均为同步表达；在诱癌的早期即可检测到较多ｃ－ｍｙｃ和Ｈａ－ｒａｓ的阳性表达细胞，而Ｋｉ－ｒａｓ的细胞数则很少，且反应强度弱；而诱癌后期，各癌基因ｍＲＮＡ阳性表达细胞均减少；至第１７周，所有癌组织内均显示三种癌基因表达阴性，或仅见个别小癌巢内少数癌细胞呈弱阳性。而癌旁肝组织内却可见三种癌基因的大量表达。结论：ｃ－ｍｙｃ和Ｈａ－ｒａｓ被激活是肝癌发生过程中的早期事件，且二者之间具有协同作用，这种协同作用在肝癌的启动上可能具有重要意义；Ｋｉ－ｒａｓ的激活则发生较晚，可能与促进细胞恶性转化有关。

C-Ha-ras-1癌基因在人宫颈癌发生中的作用

Ha-ras癌基因首先在膀胱瘤细胞系中发现,以后相继在许多肿瘤中被鉴定出来。为了探讨C-Ha-ras-1癌基因在宫颈疾患中的激活状况,我们对宫颈疾患病人宫颈活检组织DNA中C-Ha-ras-1癌基因进行了检测,现将结果报告如下。

人肺癌组织中C-Ha-ras,C-myc和C-sis基因的扩增

用同位素ａ－３２ＰｄＣＴＰ标记癌基因Ｃ－Ｈａ－ｒａｓ，Ｃ－ｍｙｃ和Ｖ－ｓｉｓ基因片段，通过点杂交和Ｓｏｕｔｈｅｒｎ杂交法检测１７例鳞癌，９例腺癌，６例其它类型肺癌组织和６例非肺癌组织中Ｃ－Ｈａ－ｒａｓ，Ｃ－ｍｙｃ和Ｃ－ｓｉｓ基因扩增的情况．结果发现：４６．８％（１５／３２）的肺癌组织有Ｃ－Ｈａ－ｒａｓ基因扩增，其中１０例（５８．８ ％）鳞癌，３例腺癌（３３．３ ％）出现Ｃ－Ｈａ－ｒａｓ基因扩增。２５％（８／３２）的肺癌组织中出现Ｃ－ｍｙｃ基因扩增，其中 ４例（２３．５％）鳞癌，３例（３３．３％）腺癌出现Ｃ－ｍｙｃ基因扩增。１８．８％（６／３２）的肺癌组织有Ｃ－ｓｉｓ基因扩增，其中４例（２３．５％）鳞癌，１例（１１．１％）腺癌出Ｃ－ｓｉｓ基因扩增。而６例非肺癌组织都役有这三种癌基因扩增。同时我们也发现有５例肺癌组织（４例鳞癌和１例腺癌）出现两种癌基因的扩增。上述结果提示：Ｃ－Ｈａ－ｒａｓ，Ｃ－ｍｙｃ和Ｃ－ｓｉｓ基因的扩增与肺癌的发生有关，特别是Ｃ－Ｈａ－ｒａｓ基因扩增在肺鳞癌的形成过程中可能起着重要作用。同时还提示：肺癌的发生与两种癌基因的扩增可能有一定关系。

大鼠心肌组织中原癌基因(c-myc,Ha-ras,c-fos)表达的随龄性改变

目的 通过研究原癌基因 (c- myc,Ha- ras,c- fos)在衰老心肌中的表达 ,探讨增龄对心肌生长相关的原癌基因的影响。方法 2 0月龄及 6月龄健康雄性 SD大鼠 ,以 Northern杂交分析 c- myc,Ha- ras,c- fos基因表达。结果 Ha- ras和 c- fos在 6月龄、2 0月龄大鼠心室肌中均表达 ,老龄鼠 c- fos的表达水平明显高于青龄鼠 (0 .52 3± 0 .0 7,0 .1 85± 0 .0 5) (P<0 .0 5) ;c- Ha- ras则无显著差异 (0 .65± 0 .0 7,0 .63± 0 .0 6) (P>0 .0 5) ;c- myc在 6月龄和 2 0月龄大鼠心室肌中均无表达。结论 原癌基因 ((c- myc,Ha- ras,c- fos等 )随增龄在心肌组织中的表达各不相同 ,探讨老年期中各基因重新激活和高表达的意义 。

c-Ha-ras、c-erbB_2癌基因产物和p53、nm23抑癌基因产物对葡萄胎恶变的预测价值

目的：探讨ｃ－Ｈａ－ｒａｓ、ｃ－ｅｒｂＢ２癌基因产物和ｐ５３、ｎｍ２３抑癌基因产物表达异常和葡萄胎恶变的关系及在预测葡萄胎恶变的价值。方法：采用针对４种基因产物的单克隆抗体，ＳＰ法免疫组织化学染色，回顾分析了≥２ａ随访证实发生恶变的葡萄胎５０例（恶变组）和未发生恶变的葡萄胎３２例（非恶变组）中４种基因产物的表达情况。结果：恶变组中ｃ－Ｈａ－ｒａｓ和ｎｍ２３基因产物的表达程度显著低于非恶变组（Ｐ＜０．０１，Ｐ＜０．００１）；恶变组中ｃ－ｅｒｂＢ２及ｐ５３基因产物的表达程度显著高于非恶变组（Ｐ＜０．０１，Ｐ＜０．０５）。进一步以４种基因产物的表达程度为变量，进行ｌｏｇｉｓｔｉｃ回归判别分析，表明４种基因产物联合应用对葡萄胎恶变的预测敏感性率为８６％，特异性率为７８．１％，而ｌｏｇｉｓｔｉｃ逐步回归判别分析显示，ｃ－ｅｒｂＢ２和ｎｍ２３基因产物联合应用时，对葡萄胎恶变的预测敏感性率为８４％，特异性率为７５％。结论：ｃ－Ｈａ－ｒａｓ、ｃ－ｅｒｂＢ２癌基因产物及ｐ５３、ｎｍ２３抑癌基因产物表达异常与葡萄胎恶变密切相关。

大鼠肝癌变过程中几种癌基因的原位表达及c-Ha-ras1点突变的研究

目的 研究大鼠肝癌变过程中几种癌基因的表达和c Ha ras 1点突变。方法 通过DEN诱发癌变启动模型和肝癌模型 ,应用免疫组化、原位杂交和MDT PCR SSCP分别观测癌基因的表达和c Ha ras1点突变。结果 c Ha ras和c myc基因的过表达参与了肝癌变的全过程 ,其中c Ha ras的过表达与癌前嗜碱性肝细胞灶、c myc基因的过表达与卵圆细胞增殖关系密切。IGF Ⅱ基因的过表达对癌前肝细胞增生灶 /结节向肝细胞癌的演进起重要作用。fms基因过表达仅与个别大鼠肝细胞癌有关。结论 大鼠肝癌变过程与c Ha ras、c myc、IGF Ⅱ和fms基因的过表达及c Ha ras1点突变有关 。

人体胃癌前病变c-Ha-ras点突变、癌基因ras产物P21表达和DNA倍体研究

本文对66例胃癌及癌前病变进行了 c-Ha-ras 第12位密码子G→T突变,ras产物P21表达及DNA 非整倍体的检测。结果表明癌基因ras 的活化和表达发生于癌变的早期,此时DNA 含量尚未明显增加,更未发生形态学上的改变。P21 表达及DNA 含量等定量指标随肠化生→非典型增生→胃癌的过程呈逐步递增趋势,提示对临床进行超早期癌诊断以及协助病理学上的疑难诊断可能具有一定意义。

互隔交链孢霉毒素诱发人胎食管上皮组织c-Ha-ras点突变的研究

体外培养的人胎儿食管上皮组织经互隔交链孢霉毒素AME或AOH短时间(4h)处理后,提取该组织高分子量DNA;同时提取未经处理的人胎儿食管上皮组织DNA(空白对照)以及食管癌组织和癌旁组织DNA。提取的高分子量DNA作为模板进行PCR反应,PCR扩增产物经HpaⅡ酶解后进行琼脂糖凝胶电泳分析结果。电泳结果显示,经HpaⅡ酶解后,空白对照组、癌组织和癌旁组织DNA的特异性扩增产物——104bp片段消失,经AME、AOH处理的人胎食管上皮组织DNA的扩增产物——104bp片段仍然存在。结果说明,AME、AOH短时间作用后,人胎食管上皮组织c-Ha-ras基因12位密码子发生了突变;Ha-ras基因突变可能是癌变过程中的“早期事件”。

p21^(Ha-ras)原核表达载体的构建、表达及纯化的研究

目的:构建PET-28a(+)-Ha-ras原核表达质粒,并表达、纯化得到p21ras蛋白,为研究p21ras在肿瘤发病中的作用和机制以及p21ras细胞内抗体抗肿瘤的研究奠定基础。方法:培养人肝癌细胞株7703,提取总RNA,通过RT-PCR特异扩增出Ha-ras cDNA,然后应用TA克隆策略将纯化后的Ha-ras插入至中间载体pMD18-T vector中得到重组质粒PMD-18T vector-ras,重组质粒PMD-18Tv ector-Ha-ras经过BamHⅠ和HindⅢ双酶切后纯化回收得到具有黏性末端的Ha-ras cDNA,将PET-28a(+)同样经过BamHⅠ和HindⅢ双酶切后纯化回收得到与ras cDNA相同的黏性末端的线形质粒片段,将具有黏性末端的ras cDNA与酶切后的PET-28a(+)定向连接,构建出重组质粒PET-28a(+)-Ha-ras,经酶切鉴定,并通过核苷酸序列测序证实。将鉴定正确的PET-28a(+)-Ha-ras转入感受态BL-21(DE3)中诱导表达,利用组氨酸"标签"(His-Tag)对p21ras进行金属螯合亲和层析纯化。结果:测序分析证实,克隆入PET-28a(+)的Ha-ras序列与Genbank登录号为"NM_005343"的Ha-ras cDNA序列一致,将重组质粒PET-28a(+)-Ha-ras转化BL21(DE3),用IPTG诱导目的蛋白表达。SDS-PAG和蛋白纯化结果表明,PET-28a(+)-Ha-Ras在E.coli中获得可溶性高表达,经Ni-NTA-树脂柱亲和层析纯化得到了PAGE纯的p21ras蛋白。结论:成功构建了原核表达载体PET-28a(+)-Ha-ras,并经表达、纯化得到活性形式的p21ras蛋白,从而为研究p21ras在肿瘤发病中的作用和机制以及P21ras细胞内抗体抗肿瘤的研究奠定了基础。