这个研究我去年（ 2010 年 2 月）在日本参加氢气医学会议期间听过 Nakao 的口头报告，确实十分有意思。当时的回忆是，通过 NFkB 信号通路实现作用，这个研究给我们现在的研究提出了重要的研究设计参考，我们过去的研究对分子机制的研究仍太肤浅，因此发表论文存在很大不足。这是非常值得我们学习的。 后来这个文章曾经以 会议 摘要 的形式发表过，现在这个文章的全文发表在 BBRC 。看来 BBRC 仍将继续关注和接受氢气相关研究，本研究来自美国匹兹堡大学 Nakao课题组 。 Hydrogen inhalation reduced epithelial apoptosis in ventilator-induced lung inju.pdf 我们最近发现呼吸氢气能减少机械通气引起的肺损伤，但这种作用的分子机制仍不清楚。因此我们希望观察呼吸氢气对 NFkB 信号通路的影响，动物模型用 C57BL6 小鼠，气管插管后，用呼吸机采用 120 次 / 分钟、 30 ml/kg 潮气量或无呼气末正压 ( 原文： tidal volume of 30 ml/kg or 10 ml/kg without positive end-expiratory pressure) ，通过呼吸机给动物呼吸含 2% 氢气的空气，对照组为 2% 氮。 NFkB 活性也就是其 DNA 结合活性采用 ESMA( 凝胶电泳迁移率转换实验 ) 和 elisa 分析。氢气能在机械通气 1 小时增加 NFkB 活性，但在机械通气 2 小时降低 NFkB 活性（ 怎么有完全相反的结果 ？）。 1 小时增加 NFkB 活性的同时抗细胞凋亡蛋白 Bcl2 和 mRNA 增加，促细胞凋亡蛋白 Bax 和 mRNA 降低。呼吸氢气能增加血氧浓度，降低肺水肿，减少炎症因子表达。 NFkB 的化学阻断剂 SN50 可以逆转这种保护效应。因此， NFkB 和 Bcl2 可能是氢气抗细胞凋亡和抗炎症作用的部分原因（假机制？）。 博主评价：本研究的亮点是采用 NFkB 将氢气的治疗作用逆转，但似乎 NFkB 本身的作用也比较复杂，将氢气的作用托付给 NFkB ，似乎不利于问题的解决，甚至将问题弄的更复杂。假如发生相反的结果， NFkB 结合活性仍有被解释的情况。作者提出本研究的不足是没有确定 NFkB 的亚型，并说正考虑其他分子途径的尝试，也说明对 NFkB 并不十分看好。因此，本研究可以作为研究思路参考，但不可以太相信。 We recently demonstrated the inhalation of hydrogen gas, a novel medical therapeutic gas, ameliorates ventilator-induced lung injury (VILI); however, the molecular mechanisms by which hydrogen amelio-rates VILI remain unclear. Therefore, we investigated whether inhaled hydrogen gas modulates the nuclear factor-kappa B (NFkB) signaling pathway. VILI was generated in male C57BL6 mice by performing a tracheostomy and placing the mice on a mechanical ventilator (tidal volume of 30 ml/kg or 10 ml/kg without positive end-expiratory pressure). The ventilator delivered either 2% nitrogen or 2% hydrogen in balanced air. NFkB activation, as indicated by NFkB DNA binding, was detected by electrophoretic mobility shift assays and enzyme-linked immunosorbent assay. Hydrogen gas inhalation increased NFkB DNA binding after 1 h of ventilation and decreased NFkB DNA binding after 2 h of ventilation, as compared with controls. The early activation of NFkB during hydrogen treatment was correlated with elevated levels of the antiapoptotic protein Bcl-2 and decreased levels of Bax. Hydrogen inhalation increased oxygen tension, decreased lung edema, and decreased the expression of proinflammatory mediators. Chemical inhibition of early NFkB activation using SN50 reversed these protective effects. NFkB activation and an associated increase in the expression of Bcl-2 may contribute, in part, to the cytoprotective effects of hydrogen against apoptotic and inflammatory signaling pathway activation during VILI.
http://www.sciencenet.cn/htmlnews/2009/8/222704.shtm 研究找到结肠癌发生中新的信号通路 由哈尔滨医科大学青年教师佟丹丹博士完成的一项课题《结肠癌中RUNX3表达及其与TGF-信号通路关系的研究》，在国内外首次从细胞实验角度证实：抑癌基因RUNX3能通过依赖TGF-和非依赖TGF-两条途径，抑制结肠癌细胞生长并诱导癌细胞死亡。该成果为人们探讨结肠癌的发病机制和指导临床治疗提供了新的认识及方向。 结直肠癌是人类常见的恶性肿瘤，除了与饮食因素有关外，还与抑癌基因失去活性、转化生长因子（TGF-）信号通路调节紊乱等多种因素相关。人类RUNX3基因于1994年发现，最初被命名为急性髓性白血病基因，以后又被称作多瘤病毒强化因子结合蛋白基因。作为一种抑癌基因，其活性减弱或消失均可导致结肠癌的发生。了解它怎样通过调节TGF-信号通路和促进结肠癌的发生，对揭示结肠癌演进奥秘十分必要。 TGF-是一种具有广泛生物学活性的细胞因子，几乎作用于所有细胞，控制细胞的一系列生命活动。一旦TGF-的信号通路发生改变，能引起多种疾病。近年来的报道表明，TGF-在人类多种肿瘤如胃肠道肿瘤、乳腺癌、卵巢癌和神经内分泌肿瘤的恶性转化、浸润及转移中均扮演重要角色，且能影响肿瘤细胞对化疗的敏感程度。RUNX3作为TGF-信号通路中的重要蛋白，参与对上皮细胞生长进行负调控。 在前期科研工作中，佟丹丹在哈医大肿瘤医院病理科主任耿敬姝教授指导下，首次发现依赖TGF-途径可抑制肿瘤细胞增殖。在此基础上，更加深入地探寻RUNX3可否通过其他途径抑制肿瘤细胞生长和增殖。期间，佟丹丹博士从结肠癌细胞系入手，深入开展实验研究，采用多种分子生物学手段，用TGF-刺激肿瘤细胞，在不同的时间点观察肿瘤细胞的生长、增殖状态，并检测TGF- 信号通路中蛋白的表达，最终成功地确定RUNX3基因不但能通过依赖TGF-途径，还可以通过不依赖TGF-两条途径发挥作用。 RUNX3 inhibits cell proliferation and induces apoptosis by TGF - beta -dependent and -independent mechanisms in human colon carcinoma cells. PMID: 19571605 Related Articles Authors: Tong, D , Jiang, Y , Li, M , Kong, D , Meng, X , Zhao, Y , Jin, Y , Bai, J , Fu, S , Geng, J Journal: Pathobiology , Vol. 76 (4): 163-9 , 2009 Abstract: BACKGROUND: Genes involved in the TGF - beta signaling pathway are often altered in several types of cancers. The TGF - beta -resistant human colon cancer cell line HT-29 has inactivated TbetaRII and deficient expression of RUNX3 and Smad4 , which are involved in the TGF - beta signaling pathway. METHODS: Western blot and immunocytochemistry were performed to confirm gene expression, the MTT assay to detect cell growth , flow cytometry to investigate the cell cycle and the TUNEL to detect cell apoptosis. RESULTS: In the absence of TGF - beta , Bim was upregulated, cell growth was inhibited and apoptosis was induced. TGF - beta treatment did not affect RUNX3 expression; however, the increase in Bim expression was significant and time dependent. Interestingly, Smad4 but not Smad2 /3 was also upregulated upon exposure to TGF - beta . This was not the case after TGF - beta treatment of parent HT-29 cells. As expected, TGF - beta further inhibited cell growth and induced apoptosis in HT-29/ RUNX3 + cells. CONCLUSION: Our data demonstrate that RUNX3 is involved in TGF - beta -dependent and -independent cell growth inhibition and apoptosis induction pathways. Affiliation: Department of Pathology, Tumor Hospital, Harbin Medical University, China . 相关文献： Title: Molecular pathology of RUNX3 in human carcinogenesis. PMID: 19682550 Related Articles Authors: Subramaniam, M M , Chan, J Y , Yeoh, K G , Quek, T , Ito, K , Salto-Tellez, M Journal: Biochim Biophys Acta , 2009 Abstract: A major goal of molecular biology is to elucidate the mechanisms underlying cancer development and progression in order to achieve early detection, better diagnosis and staging and novel preventive and therapeutic strategies. We feel that an understanding of Runt-related transcription factor 3 ( RUNX3 )-regulated biological pathways will directly impact our knowledge of these areas of human carcinogenesis. The RUNX3 transcription factor is a downstream effector of the transforming growth factor - beta ( TGF - beta ) signaling pathway, and has a critical role in the regulation of cell proliferation and cell death by apoptosis, and in angiogenesis, cell adhesion and invasion. We previously identified RUNX3 as a major gastric tumor suppressor by establishing a causal relationship between loss of function and gastric carcinogenesis. More recently, we showed that RUNX3 functions as a bona fide initiator of colonic carcinogenesis by linking the Wnt oncogenic and TGF - beta tumor suppressive pathways. Apart from gastric and colorectal cancers, a multitude of epithelial cancers exhibit inactivation of RUNX3 , thereby making it a putative tumor suppressor in human neoplasia. This review highlights our current understanding of the molecular mechanisms of RUNX3 inactivation in the context of cancer development and progression. Affiliation: Cancer Science Institute of Singapore (CSI), National University of Singapore , Singapore . Title: Epigenetic inactivation of RUNX3 in microsatellite unstable sporadic colon cancers. PMID: 15386381 Related Articles Authors: Goel, A S , Arnold, C N , Tassone, P F , Chang, D K , Niedzwiecki, D , Dowell, J M , Wasserman, L , Compton, C , Mayer, R J , Bertagnolli, M M , Boland, C R Journal: Int J Cancer , Vol. 112 (5): 754-9 , 2004 Abstract: Runt domain transcription factors are important targets of TGF - beta superfamily proteins and play a crucial role in mammalian development. Three mammalian runt-related genes, RUNX1 , RUNX2 and RUNX3 , have been described. RUNX3 has been shown to be a putative tumor suppressor gene localized to chromosome 1p36, a region showing frequent loss of heterozygosity events in colon, gastric, breast and ovarian cancers. Because of the important role of TGF - beta signaling in the human colon, we hypothesized that RUNX3 may serve as a key tumor suppressor in human colon cancers and colon cancer -derived cell lines. We examined RUNX3 expression and the frequency of RUNX3 promoter hypermethylation in 17 colon cancer cell lines and 91 sporadic colorectal cancers. Semiquantitative analysis of RUNX3 transcripts was performed by RT-PCR and de novo methylation of the RUNX3 promoter was studied by a methylation-specific PCR (MSP) assay. Nineteen of 91 informative tumors (21%) and 11 of 17 (65%) colon cancer cell lines exhibited hypermethylation of the RUNX3 promoter. Interestingly, RUNX3 promoter hypermethylation was more common in tumors exhibiting high frequency of microsatellite instability (MSI-H) (33% of MSI-H vs. 12% of MSI-L/MSS tumors; p = 0.012). Hypermethylation of the RUNX3 promoter correlated with loss of mRNA transcripts in all cell lines. RUNX3 promoter methylation was reversed and its expression restored in SW48 and HCT15 colon cancer cells after treatment with the demethylating agent 5-aza-2'-deoxycytidine, indicating that loss of expression is caused by epigenetic inactivation in colon carcinogenesis. This is the first demonstration of frequent de novo hypermethylation of the RUNX3 promoter in sporadic colon cancers. The significant association of RUNX3 promoter hypermethylation with MSI -H colon cancers suggests that RUNX3 is a novel target of methylation, along with the hMLH1 gene, in the evolution of MSI-H colorectal cancers. Affiliation: Department of Medicine and Comprehensive Cancer Center, University of California San Diego , La Jolla, CA , USA . Pubmed MeSH: Core Binding Factor Alpha 3 Subunit , DNA Methylation , DNA-Binding Proteins , Humans , Tumor Cells, Cultured 信息分析平台： http://www.gopubmed.org/web/gopubmed/1?WEB0caljf8evalkeI1rI1I00d000j10040001rl 检索策略： colonic cancer and RUNX3 and TGF- 相关文献信息分析结果： Top Years Publications 2004 21 2005 18 2007 10 2003 9 2009 8 2002 6 2000 6 1999 5 2006 4 1998 4 2001 3 2008 3 1997 3 Top Countries Publications USA 39 Japan 18 Germany 7 United Kingdom 7 South Korea 6 Australia 4 China 3 Italy 2 Singapore 2 Spain 2 France 2 Hong Kong S.A.R., China 1 Norway 1 Ireland 1 Portugal 1 Finland 1 Sweden 1 Croatia 1 Netherlands 1 1 2 3 Top Cities Publications Baltimore 12 Seoul 5 San Diego 5 Boston 5 Tokyo 5 Sapporo 4 Regensburg 3 Dallas 3 Houston 3 Heidelberg 2 Washington 2 Tochigi 2 Singapur 2 Oxford 2 Nagoya 2 London 2 Santa Monica 2 Barcelona 2 Paris 2 Bologna 1 1 2 3 1 2 3 Top Journals Publications Cancer Res 12 Int J Cancer 11 Oncogene 9 Clin Cancer Res 6 Cancer Biol Ther 5 Gastroenterology 4 Fam Cancer 2 Hum Pathol 2 Mol Cancer 2 Carcinogenesis 2 Brit J Cancer 2 Neoplasia 2 Oncology 2 Bmc Cancer 2 Plos One 2 Gene Chromosome Canc 1 Clin Gastroenterol Hepatol 1 Am J Gastroenterol 1 J Cancer Res Clin 1 Mol Carcinogen 1 1 2 3 1 2 3 ... 32 Top Authors Publications Boland C 6 Goel A 6 Meltzer S 6 Issa J 6 Arnold C 5 Yin J 5 Baylin S 5 Niedzwiecki D 4 Compton C 4 Mayer R 4 Bertagnolli M 4 Wang S 4 Abraham J 4 Herman J 4 Ogino S 4 Kawasaki T 4 Imai K 4 Toyota M 4 Hamelin R 4 Sato F 4 1 2 3 ... 32 1 2 3 ... 43 Top Terms Publications Colorectal Neoplasms 100 Humans 92 Genes 82 Neoplasms 77 Methylation 73 DNA Methylation 73 methylation 72 Microsatellite Repeats 64 Microsatellite Instability 55 Mutation 53 Colonic Neoplasms 51 DNA 47 Cell Line 44 Polymerase Chain Reaction 43 Carcinoma 40 Proteins 39 Patients 38 Tissues 33 regulation of cell cycle 28 Nuclear Proteins 28 1 2 3 ... 43 Top Terms Publications regulation of cell cycle 27 Cell Line, Tumor 26 Gene Expression Regulation, Neoplastic 25 Middle Aged 25 Neoplasm Proteins 24 DNA-Binding Proteins 24 RNA, Messenger 23 Aged 22 Genes, Tumor Suppressor 22 Reverse Transcriptase Polymerase Chain Reaction 22 positive regulation of mismatch repair 20 negative regulation of mismatch repair 20 regulation of mismatch repair 20 mismatch repair complex 20 DNA Mismatch Repair 20 mismatch repair 20 Nuclear Proteins 20 Carrier Proteins 20 DNA, Neoplasm 20 Loss of Heterozygosity 19 1 2 3 4 ... 50 Top Terms Publications Adult 19 Genes, Suppressor 19 regulation of gene expression 19 Adaptor Proteins, Signal Transducing 18 gene expression 18 Base Sequence 18 DNA methylation 16 Adenocarcinoma 16 Genomics 15 Genome 15 Immunohistochemistry 15 Mucous Membrane 15 Transcription Factors 15 chromosome 15 gene silencing 14 Exons 13 CpG Islands 13 Stomach Neoplasms 13 Gene Expression 13 Alleles 12 1 2 3 4 5 ... 50
标签: 信号通路
