Functional Analysis of Tumor Suppressor ING Family in Oral Cancer

抑癌基因ING家族在口腔癌中的功能分析

• 批准号: 17591911
• 负责人: MEHMET Gunduz
• 金额: $ 2.18万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591911/
• 关键词: Oral cancer; ING family; Splicing variant; p53; Apoptosis; Tumor suppressor; Methylation

1. Oral cancer occurs through inactivation of multiple tumor suppressor genes and activation of oncogenes. In this research, we examined the functions of the ING family members genes ING1b, ING1c and ING3, which we previously identified. mRNA expression analysis of splicing variants ING1b and ING1c showed 30% and 58% decrease in tumor samples as compared with the matched normal counterparts. On the other hand, ING3 mRNA was down-regulated in 50% of cancer samples. Restriction endonuclease-based methylation analysis demonstrated 51% methylation of ING1b variant, while it was not detected in ING1c. These results suggested that different splicing variants and members have various role and mechanisms in oral cancer.2. Flag-tagged expression vectors of ING1b, ING1c and ING3 were constructed for in vitro functional analysis. Each of these vectors alone and combination with p53 expression vectors were transfected into the cell lines of HEK293, SCCKN, HSC3, HSC3 and cell proliferation and apop … More tosis assay were performed. These assays showed that INGlc-p53 combination gave a better cell inhibitory effect. Each gene and combination with p53 also resulted in apoptotic changes.3. To examine the response of the ING family and proapoptotic as well as antiapoptotic genes to the cytotoxic drugs, we treated the cells with 5-FU, Cisplatin and Etoposide and mRNA expressions of ING1lb, ING1c, ING3, Bax and Bc1-2 were checked by quantitative RT-PCR. These experiments showed different reactions with various drugs in each cell line, suggesting that signaling of each variant and member has different mechanism.4. We also examined ING3 mRNA expression in over 70 cancer samples and compared with clinicopathological variables. This study demonstrated ING3 mRNA value as a possible prognostic marker. The samples with ING3 down-regulation had bad prognosis with a 40% survival rate as compared to 60% survival in normal ING3 mRNA expression. Future studies will include suppression work of each ING family member by siRNA and examination of the results in animal experiments. Identification of all networks of ING family will lead to development of novel molecular therapies In various cancer types including oral cancer. Less

1.口腔癌是通过多种肿瘤抑制基因的失活和癌基因的激活而发生的。在这项研究中，我们研究了ING家族成员基因ING 1b，ING 1c和ING 3的功能，我们以前确定。剪接变体ING 1b和ING 1c的mRNA表达分析显示，与匹配的正常对照相比，肿瘤样品中的ING 1b和ING 1c分别降低了30%和58%。另一方面，ING 3 mRNA在50%的癌症样本中下调。基于限制性内切酶的甲基化分析显示ING 1b变异体的甲基化率为51%，而ING 1c中未检测到甲基化。这些结果提示不同的剪接变异体和成员在口腔癌中具有不同的作用和机制.构建了带有Flag标签的ING 1b、ING 1c和ING 3表达载体，用于体外功能分析。将这些载体单独或与p53表达载体组合转染入HEK 293、SCCKN、HSC 3、HSC 3细胞系中，细胞增殖和apo ...更多信息 进行Tosis测定。这些测定显示INGlc-p53组合产生更好的细胞抑制作用。各基因及与p53基因联合也可导致细胞凋亡的改变.为了检测ING家族和促凋亡基因以及抗凋亡基因对细胞毒性药物的反应，我们用5-FU、顺铂和足叶乙甙处理细胞，并通过定量RT-PCR检测ING 11b、ING 1c、ING 3、Bax和Bc 1 -2的mRNA表达。这些实验显示不同的药物在不同的细胞系中有不同的反应，表明每个变体和成员的信号传导具有不同的机制.我们还检测了70多个癌症样本中的ING 3 mRNA表达，并与临床病理变量进行了比较。这项研究表明ING 3 mRNA值作为一个可能的预后标志物。ING 3表达下调的患者预后差，存活率为40%，而ING 3 mRNA表达正常的患者存活率为60%。未来的研究将包括通过siRNA对每个ING家族成员的抑制工作以及在动物实验中检查结果。ING家族所有网络的鉴定将导致在包括口腔癌在内的各种癌症类型中开发新的分子疗法。少

项目成果

Palatinose Based Enteral Formula (MHN-01) Carbohydrate-adjusted Fluid Diet in Improving the Metabolism of Carbohydrates and Lipids in Patients with Esophageal Cancer Complicated by Diabetes Mellitus.

帕拉金糖肠内配方（MHN-01）碳水化合物调整流质饮食可改善合并糖尿病的食道癌患者碳水化合物和脂质的代谢。

• 期刊: J Surg Res 2007 Jan 23 [Epub ahead of print]
• 作者: Fujiwara T;Naomoto Y;Motoki T;Shigemitsu K;Shirakawa Y;Yamatsuji T;Kataoka M;Haisa M;Fujiwara T;Egi M;Morimatsu H;Hanazaki M;Katayama H;Morita K;Mizumoto K;Asou T;Arima H;Sasaki H;Matsuura M;Gunduz M;Tanaka N. Effects of a Novel
• 通讯作者: Tanaka N. Effects of a Novel

Chapter 13 page : 219-230, In Real Time PCR, Ed. M.Tevfik Dorak, Taylor ＆ Francis, London 2006

第 13 章页：219-230，实时 PCR，M.Tevfik Dorak，Taylor & Francis，伦敦，2006 年

• 发表时间: 2006
• 作者: Cakilci B;Gunduz M
• 通讯作者: Gunduz M

Heparanase Promotes Angiogenesis through Cox-2 and HIF1α.

乙酰肝素酶通过 Cox-2 和 HIF1α 促进血管生成。

• 发表时间: 2007
• 期刊: Med Hypotheses 68(1)
• 作者: Naomoto Y;Gunduz M;Takaoka M;Okawa T;Gunduz E;Nobuhisa T;Kobayashi M;Shirakawa Y;Yamatsuji T;Sonoda R;Matsuoka J;Tanaka N.
• 通讯作者: Tanaka N.

Nuclear interaction of EGFR and STAT3 in the activation of iNOS/NO pathway

EGFR 和 STAT3 的核相互作用在 iNOS/NO 通路激活中的作用

• 发表时间: 2005
• 期刊: Cancer Cell 7(6)
• 作者: Nobuhisa T;Naomoto Y;Ohkawa T;Takaoka M;Ono R;Murata T;Gunduz M;Shirakawa Y;Yamatsuji T;Haisa M;Matsuoka J;Tsujigiwa H;Nagatsuka H;Nakajima M;Tanaka N.;Okawa T;Lo HW
• 通讯作者: Lo HW

Carbohydrate-adjusted Fluid Diet in Improving the Metabolism of Carbohydrates and Lipids in Patients with Esophageal Cancer Complicated by Diabetes Mellitus

碳水化合物调整流质饮食改善食管癌合并糖尿病患者碳水化合物和脂质代谢

• 期刊: J Srg res (In press)
• 作者: Fujiwara T;Naomoto Y;Motoki T;Shigemitsu K;Shirakawa Y;Yamatsuji T;Kataoka M;Haisa M;Fujiwara T;Egi M;Morimatsu H;Hanazaki M;Katayama H;Morita K;Mizumoto K;Asou T;Arima H;Sasaki H;Matsuura M;Gunduz M;Tanaka N. Effects of a Novel;Fujiwara T
• 通讯作者: Fujiwara T