New syngeneic inflammatory‐related lung cancer metastatic model harboring double KRAS/WWOX alterations

New mouse models with specific drivers of genetic alterations are needed for preclinical studies. Herein, we created and characterized at the genetic level a new syngeneic model for lung cancer and metastasis in Balb‐c mice. Tumor cell lines were obtained from a silica‐mediated airway chronic inflammation that promotes tumorigenesis when combined with low doses of N‐nitrosodimethylamine, a tobacco smoke carcinogen. Orthotopic transplantation of these cells induced lung adenocarcinomas, and their intracardiac injection led to prominent colonization of various organs (bone, lung, liver and brain). Driver gene alterations included a mutation in the codon 12 of KRAS (G–A transition), accompanied by a homozygous deletion of the WW domain‐containing oxidoreductase (WWOX) gene. The mutant form of WWOX lacked exons 5–8 and displayed reduced protein expression level and activity. WWOX gene restoration decreased the in vitro and in vivo tumorigenicity, confirming the tumor suppressor function of this gene in this particular model. Interestingly, we found that cells displayed remarkable sphere formation ability with expression of specific lung cancer stem cell markers. Study of non‐small‐cell lung cancer patient cohorts demonstrated a deletion of WWOX in 30% of cases, with significant reduction in protein levels as compared to normal tissues. Overall, our new syngeneic mouse model provides a most valuable tool to study lung cancer metastasis in balb‐c mice background and highlights the importance of WWOX deletion in lung carcinogenesis.     

Investigation of osteosarcoma genomics and its impact on targeted therapy:an international collaboration to conquer human osteosarcoma

Osteosarcoma is a genetical y unstable malignancy that most frequently occurs in children and young adults. The lack of progress in managing this devastating disease in the clinic has prompted internat...     

WWOX蛋白在非小细胞肺癌中的表达及临床意义

目的通过WWOX蛋白在非小细胞肺癌（NSCLC）癌灶与癌旁正常组织中表达的比较,探讨WWOX蛋白在NSCLC发生、发展中的作用。方法用免疫组化elivision法检N51例NSCLC组织标本中WWOX蛋白的表达,与20例癌旁正常组织标本作对照,并分析WWOX蛋白与NSCLC患者年龄、性别、组织分化程度、病理类型、吸烟及淋巴结转移等关系。结果WWOX蛋白在NSCLC组织中表达的阳性率25．49％（13／51）,癌旁正常组织中表达的阳性率85．0％（17／20）,二者比较差异有统计学意义（P〈0．01）。WWOX蛋白阳性率与NSCLC组织分化程度、病理类型、吸烟、有无淋巴结转移有相关关系,而与患者的年龄、性别及肿瘤TNM分期无相关性。结论WWOX蛋自在NSCLC组织中表达明显低于癌旁正常组织,WWOX蛋白的表达与NSCLC的多种重要临床病理预后因素相关,有望成为判断NSCLC临床发展的分子标记。     

抑癌基因WWOX及其与恶性肿瘤的研究进展

WWOX(WW domain containing oxidoreductase)基因是2000年由Bednarek等[1]在染色体普通脆性位点(common frigal of sites,CFSs) FRA16D(16q23～32q24.1)区域克隆出的新基因.     

非小细胞肺癌中WWOX蛋白及ErbB2蛋白的联合检测

 目的 探讨WWOX(WW domain-containing oxidoreductase)基因在非小细胞肺癌组织中的表达及与ErbB2蛋白之间的关系。 方法 用免疫组化方法检测50例肺鳞癌,31例肺腺癌及20例癌旁正常组织中WWOX基因蛋白的表达,并分析其与各临床病理指标及ErbB2蛋白之间的相关性。 结果 WWOX基因在72.8%的非小细胞肺癌中失表达或表达减少,而在癌旁正常组织中有80.0%正常表达。WWOX基因的表达与组织类型、细胞分化程度密切相关（P<0.05）,在鳞癌和低分化癌中WWOX基因失表达或表达减少的发生率更高。肺癌中ErbB2的过度表达与肺癌的临床分期、淋巴结转移相关,WWOX蛋白阳性表达与ErbB2负相关(r=–0.239,Ｐ<0.05)。 结论 WWOX蛋白的低表达与ErbB2的过表达可以协同判断预后较差的非小细胞肺癌。     

Microarray analysis reveals down-regulation of the tumour suppressor gene WWOX and up-regulation of the oncogene TYMS in intracranial sporadic meningiomas

BACKGROUND: In order to investigate pathways that may influence on tumour development in meningiomas, we performed high throughput microarray analysis of the RNA expression and DNA copy number of 22 WHO grade I and five WHO grade II meningiomas. Since meningiomas derive from arachnoid cap cells, we used samples from four patients operated for arachnoid cysts as control tissue. RESULTS: The expression of the tumour suppressor gene WW containing oxidoreductase (WWOX) was down-regulated, and the thymidylate synthase (TYMS) oncogene was up-regulated in all meningiomas as compared to arachnoid cysts. Unsupervised RNA cluster analysis showed that fibrous meningiomas gathered in two clusters, and thus were more homogeneous than the other meningiomas. The other histological subgroups could not be linked to any uniform gene expression signatures. Rearrangements were most abundant on chromosomes 1 and 22, but were identified on all except chromosome 16. The fibrous and mixed meningiomas generally had chromosomal deletions. Duplications were more frequent in the meningothelial meningiomas. WHO grade II meningiomas had increased chromosomal instability. CONCLUSION: Decreased expression of the WWOX tumour suppressor gene and increased expression of the TYMS oncogene may be of importance for the development of human intracranial meningiomas. We have identified several genes (BMPR1B, DMD, RAMP1) with expression signatures specific for fibrous meningiomas. CGH analysis revealed distinct chromosomal patterns in relation to the histological subtypes of the meningiomas.     

尼古丁对人牙龈成纤维细胞中抑癌基因WWOX表达的影响

目的观测不同浓度尼古丁培养液对体外培养的人牙龈成纤维细胞（hmu-na gingival fibroblast,HGF）内抑癌基因WWOX转录水平的影响,从分子水平探讨吸烟对口腔肿瘤发生的影响。方法取第5代HGF,以FBS作为空白对照组,实验组分别加入终浓度为0.01、0.1、1μg/mL的尼古丁,标准条件下培养4天,提取细胞总RNA进行RT-PCR反应,测量基因转录水平。结果WWOX基因mRNA在空白组的比值为0.89 720,加入尼古丁培养液后HGF中WWOX基因mR-NA表达水平降低,低浓度组为0.88 650,中浓度组为0.75 320,高浓度组为0.61 760。结论尼古丁影响了HGF中WWOXmRNA的转录,随着尼古丁浓度的增加WWOXmRNA合转录水平降低。     

WWOX与P53在食管鳞癌中的表达及其临床意义

目的分析WWOX与P53蛋白在食管鳞癌中的表达,探讨它们与食管鳞癌临床病理特征的关系。方法用免疫组化法检测40例正常食管黏膜、癌旁食管黏膜及癌组织标本的WWOX与P53蛋白产物的表达情况,并结合临床病理特征进行分析。结果在食管鳞癌组织中,中分化组与低分化组WWOX蛋白阳性表达率、P53蛋白阳性表达率差异均有统计学意义(P<0.05);II期与III~IV期间WWOX蛋白异常表达差异有统计学意义(P<0.05),I期或II期均与III~IV期间P53蛋白异常表达差异有统计学意义(P<0.05);有、无淋巴结转移WWOX蛋白阳性表达率、P53蛋白阳性表达率差异均有统计学意义(P<0.05)。食管鳞癌组织中,WWOX蛋白阴性表达与P53蛋白阳性表达呈正相关(R s=0.514,P<0.001)。结论WWOX蛋白与P53蛋白异常表达与食管鳞癌的癌组织分化程度、临床分期及转移有关。在食管鳞癌中WWOX蛋白与P53蛋白表达密切相关。联合检测WWOX蛋白与P53蛋白,可以为食管鳞癌的诊断、治疗及判断预后提供有意义的生物学指标。     

WWOX基因在贲门腺癌中的异常甲基化及其表达

目的:探讨贲门腺癌(gastric cardia adenocarcinoma,GCA)中WWOX( WW domain-containing oxidoreductase)基因启动子区及第1外显子区的甲基化状态及其对表达的影响,并分析贲门腺癌中WWOX与Smad4(mothers against decapentaplegic homolog 4)蛋白表达之间的关系.方法:分别应用甲基化特异性PCR和免疫组织化学法检测贲门腺癌组织及其相应癌旁组织中WWOX基因的甲基化和蛋白的表达情况:应用免疫组织化学法检测贲门腺癌组织及其相应癌旁组织中Smad4蛋白的表达.结果:贲门腺癌组织中WWOX基因启动子区(- 382～- 41 bp)及第1外显子区(- 27～+344 bp)的甲基化率[26.4％ (33/125)和23.2％ (29/125)]显著高于其相应的癌旁常组织(0％和0％)(P＜0.01),并与贲门腺癌TNM分期和上消化系统肿瘤家族史有关(P＜0.05).贲门腺癌组织中WWOX蛋白的表达阳性率显著低于相应癌旁组织(P＜0.01),WWOX蛋白的表达与WWOX启动子区和第1外显子区甲基化状态之间呈负相关(r=-0.216,r=-0.255,P＜0.05).贲门腺癌组织中Smad4蛋白表达的阳性率(47.2％)显著低于相应癌旁组织(94.4％)(P＜0.01),且与WWOX在贲门腺癌中的表达呈明显的正相关(r=0.622,P＜0.01).结论:WWOX基因启动子区及第1外显子区的高甲基化导致的基因表达下调可能在贲门腺癌中发挥重要作用.