Study on the role of DNA methyltransferases in regulation of the expression of genes associated with the control of cell cycle

DNA甲基转移酶在细胞周期控制相关基因表达调控中的作用研究

• 批准号: 14571880
• 负责人: BUKAWA Hiroki
• 金额: $ 2.56万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571880/
• 关键词: DNA methyltransferase; oral squamous cell carcinoma; DNA methylation; APC gene; p16 gene; FEZ1 gene; KAI1 gene; survivin; 口腔扁平上皮癌; DNA methyltransferase; APC遺伝子; FEZ1遺伝子; p73遺伝子

To estimate the involvement of methylation in the down-regulation of the genes, APC, p16, FEZ1, KAI1, and survivine, wehich were associated with the regulation of cell cycle, we examined the expression revels of them and methyltransferases, DNMT1, DNMT3A, and DNMT3B, which were believed to be associated with carcinogenesis, in 8 cell lines, SAS, HSC-2, HSC-3, HSC-4, Ca9-22, OK-92, HO-1-u-1, and HO-1-N-1, which were derived from oral carcinoma, and clinical tissue samples of oral cancer. Additionally, restoration of the gene expression by 5-Aza-C, one of the demethylation agents, was examined. The hyper methylation of CDKN2A/p16 gene was detected in 24 (48.0%) of 50 oral squamous cell carcinoma cases (OSCC) and the restoration by 5-Aza-C was found in 6 (75.0%) of 8 cell lines. The down expression of APCgene was detected in 15 (30.0%) of 50cases, and hypermethylation of CPG island was found 12 (24.0%) of the all cases. Suppressed expression and hypermethylation of FEZ1gene was detected i … More n 11 (35.5%) and in 8 (25.8%) of 31 case, respectively. Out of the 31 cases, hypermethylation was detected and restoration of the gene expression by 5-Aza-C was found in all of the 8 cell lines. Silence or suppressed expression of KAI1 gene was detected in 84 of 101 OSCC cases. However, neither hypermethylation of CPG island of the promoter region nor restoration of the gene expression by 5-Aza-C was not found. The over-expression of surviving gene was detected in 58% of OSCC cases and in 37% of precancerous lesion, leukoplakia. On the other hand, the expression of surviving gene was not detected by RT-PCRin all of the 9 normal oral epitherium tissues and DNA methylation was confirmed by methylation specific PCR in 4 of the 9 normal specimens and the gene expression was restored by 5-Aza-Ctreatment in 2 of 8 cell lines. The over-expression of DNA methyltransferase, DNMTs ; DNMT1, DNMT3A, and DNMT3B, which were believed to be associated with carcinogenesis, were high frequently detected in OSCC. The rates of the expression of DNMTI, DNMT3A, and DNMT3B in OSCC were 72%, 56%, and 64%, respectively. There was no significant correlation between the expression levels and the situations of hypermethylation of the tumor suppressor and oncogenes examined and the expression levels of methyltransferases. Our data suggest that hypermethylation of the gene may be one of the major mechanisms for inactivation of the genes in oral carcinogenesis and that besides methyltransferases, some transcription factors involved in the suppression of gene expression might play an important roles in ora oncogenesis. Less

为了评估甲基化在与细胞周期调控相关的基因APC、p16、FEZ 1、KAI 1和生存素下调中的作用，我们检测了它们以及被认为与癌发生相关的甲基转移酶DNMT 1、DNMT 3A和DNMT 3B在8种细胞系SAS、HSC-2、HSC-3、HSC-4、Ca 9 -22、OK-92、HO-1-u-1和HO-1-N-1，它们来源于口腔癌和口腔癌的临床组织样品。此外，通过5-Aza-C（去甲基化剂之一）检测基因表达的恢复。50例口腔鳞癌中24例（48.0%）存在CDKN 2A/p16基因甲基化，8株细胞系中6株（75.0%）经5-Aza-C处理后恢复甲基化。APC基因表达下调15例（30.0%），CPG岛甲基化12例（24.0%）。FEZ 1基因表达抑制和甲基化， ...更多信息 11例（35.5%），8例（25.8%）。在31例病例中，检测到高甲基化，并在所有8个细胞系中发现5-Aza-C恢复基因表达。101例口腔鳞癌中有84例KAI 1基因表达被抑制或沉默。58%的OSCC和37%的癌前病变白斑中存在Survivin基因的过度表达，而5-Aza-C对Survivin基因表达的恢复作用不明显。9例正常口腔上皮组织中RT-PCR均未检测到Survivin基因的表达，甲基化特异性PCR检测到4例正常口腔上皮组织中存在DNA甲基化，经5-Aza-C处理后，2例正常口腔上皮组织中Survivin基因表达恢复。DNA甲基转移酶（DNMTs）、DNMT 1、DNMT 3A和DNMT 3B在口腔鳞癌中的高表达被认为与口腔鳞癌的发生有关。DNMT 1、DNMT 3A和DNMT 3B在口腔鳞癌中的表达率分别为72%、56%和64%。抑癌基因和癌基因的甲基化程度与甲基转移酶的表达水平无明显相关性。我们的数据表明，该基因的高甲基化可能是口腔癌发生的主要机制之一的基因失活，除了甲基转移酶，一些转录因子参与抑制基因表达可能在口腔癌的发生中发挥重要作用。少

项目成果

Uzawa K, Ono K, Suzuki H, Yokoe H, Tanzawa H.: "High prevalence of decreased expression of KAI1 metastasis suppressor in human oral carcinogenesis"Clinical Cancer Research. 8. 828-835 (2002)

Uzawa K、Ono K、Suzuki H、Yokoe H、Tanzawa H.：“人类口腔癌发生过程中 KAI1 转移抑制因子表达降低的高发生率”临床癌症研究。

Uzawa K, Ono K, Suzuki H, Tanaka C, Yakushiji T, Yamamoto N, Yokoe H, Tanzawa. H.: "High prevalence of decreased expression of KAI1 metastasis suppressor in human oral carcinogenesis."Clinical Cancer Research. 8. 828-835 (2002)

宇泽 K、小野 K、铃木 H、田中 C、药师寺 T、山本 N、横江 H、丹泽。

Uzawa K, Ono K, Suzuki H, Tanaka C, Yakushiji T, Yamamoto N, Yokoe H, Tanzawa H.: "High prevalence of decreased expression of KAI1 metastasis suppressor in human oral carcinogenesis."Clinical Cancer Research. 8. 828-835 (2002)

Uzawa K、Ono K、Suzuki H、Tanaka C、Yakushiji T、Yamamoto N、Yokoe H、Tanzawa H.：“人类口腔癌发生过程中 KAI1 转移抑制因子表达降低的发生率很高。”临床癌症研究。

Ono K, Uzawa K, Nakatsuru M, Shiiba M, Mochiba Y, Tada A, Bukawa H, Miyakawa A, Yokoe H, Tanzawa H.: "Down-rogulation of FEZ1/LZTS1 gene with frequent loss of heterozygosity in oral squamous cell carcinomas."Int J Oncol. 23. 297-302 (2003)

Ono K、Uzawa K、Nakatsuru M、Shiiba M、Mochiba Y、Tada A、Bukawa H、Miyakawa A、Yokoe H、Tanzawa H.：“口腔鳞状细胞癌中 FEZ1/LZTS1 基因下调并频繁丢失杂合性

Tanaka C, Uzawa K, Shibahara T, Yokoe H, Noma H, Tanzawa H.: "Expression of an inhibitor of apoptosis, surviving, in oral carcinogenesis."J Dent Res. 82. 607-611 (2002)

Tanaka C、Uzawa K、Shibahara T、Yokoe H、Noma H、Tanzawa H.：“在口腔癌发生中存活的细胞凋亡抑制剂的表达。”J Dent Res。