Inhibitory effects on tumor invasion and metastasis induced by transfection of anti-sense E1AF,an ets-oncogene family transcription factor.

ets-癌基因家族转录因子反义E1AF转染对肿瘤侵袭和转移的抑制作用。

• 批准号: 08672063
• 负责人: SHINDOH Masanobu
• 金额: $ 1.41万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08672063/
• 关键词: E1AF; ets-oncogene; antisense; tumor invasion; p21^<waf1; cip1>; ets-omcgene; timor innasion; cip1>; EIAF; Matrix metalloproteinase; Oral Squamows cell carvinoma; imrasion; anti sense vector

E1AF is a newly identified human ets-family transcription factor. We have reported that E1AF can up-requlate transcription of matrix metalloproteinase (MMP) genes and confers invasive phenotype on human cancer cells. HSC3 is an oral squamous cell carcinoma-derived cell line, and it manifests high levels of E1AF,and MMP-1 and -9 gene expression that are associated with invasive potential. We reconstructed an E1AF antisense expression vector, transfected HSC3 cells with the vector and obtained HSC3AS cells which express E1AF antisense RNA.HSC3AS showed decreasing m, -3 and -9. moreover, HSC3AS showed lower invasive potential in in vitro three-dimensional raft culture and in vivo implantation into nude mice. These results imply that transfection of antisense E1AF inhibits tumor invasion by down-regulating MMP genes.Thus, E1AF is thought to be highly correlated with malignant potentials of cancer cells, however, little is known about E1AF expression and cancer cell malignancies in in vivo … More tumors. We examined 27 oral SCC speciment using RT-PCR,Southern blot hybtidization and in situ hybridization (ISH) and compared to the clinico-pathological parameters. Among the 27 patients, E1AF was detected in 15 cases. E1AF mRAN was detected in 13 of 17 invasive SCC_s, whereas the majority of SCC_s not expressing E1AF showed an expansive growth pattern. Increased prevalence of E1AF-positive oral SCC was observed in cases with nodal metastasis. These results indicate that E1AF may be involved in cancer cell malignancies through its ability to promote invasive potentialp21^<Waf1/Cip1> is one of the key requlatory proteins in cell cycle, terminal differentiation and apoptosis. its promoter was shown to be transactivated by the wild-type p53 protein as well as in a p53-independent manner. We demonstrate that E1AF,an ets-related transcription factor, activates the human p21^<Waf1/Cip1> promoter by interacting with the ets-binding sites located close to the two previously identified p53-responsive elements. Northern blot analysis revealed that p21^<Waf1/Cip1> and E1AF were correlatively uprequlated in response to cisplatin treatment in SiHa cells. Transient expression assays demonstrated that E1AF can activate the p21^<Waf1/Cip1> promoter-driven luciferase reporter gene in SiHa cells. The p21^<Waf1/Cip1> promoter activity was also increased in p53-null Saos2 osteosarcoma cells, but was markedly reduced when the etsbinding sites were deleted. These results indicate that E1AF positively requlates transcription from the p21^<Waf1/Cip1> promoter in response to genotoxic stresses. Less

E1 AF是一个新发现的人类ets家族转录因子。我们已经报道了E1 AF可以上调基质金属蛋白酶（MMP）基因的转录，并赋予人癌细胞侵袭表型。HSC 3是一种口腔鳞状细胞癌衍生的细胞系，它表现出高水平的E1 AF，MMP-1和MMP-9基因表达，这些基因表达与侵袭潜力相关。构建E1 AF反义表达载体，转染HSC 3细胞，获得表达E1 AF反义RNA的HSC 3AS细胞，HSC 3AS细胞的m值、-3和-9均降低。此外，HSC 3AS在体外三维筏培养和裸鼠体内植入中显示出较低的侵袭潜力。这些结果表明，反义E1 AF基因转染通过下调MMP基因抑制肿瘤侵袭，因此，E1 AF被认为与肿瘤细胞的恶性潜能密切相关，但E1 AF基因表达与肿瘤细胞恶性程度的关系在体内研究中还知之甚少 ...更多信息 肿瘤的应用RT-PCR、Southern杂交和原位杂交技术对27例口腔鳞癌标本进行检测，并与临床病理参数进行比较。27例患者中，15例检出E1 AF。17例浸润性SCC中13例表达E1 AF mRAN，而大多数不表达E1 AF的SCC呈扩张性生长。在有淋巴结转移的病例中，观察到E1 AF阳性口腔鳞癌的患病率增加。这些结果表明E1 AF可能通过促进细胞的侵袭能力而参与恶性肿瘤的发生。p21 <Waf 1/Cip 1>是细胞周期、终末分化和凋亡的关键调控蛋白之一。其启动子被野生型p53蛋白反式激活，并且以p53非依赖性方式激活。我们证明，E1 AF，一个ets相关的转录因子，激活人类p21^<Waf 1/Cip 1>启动子通过与ets结合位点的相互作用位于接近两个先前确定的p53响应元件。北方印迹分析显示顺铂处理后SiHa细胞中p21^<Waf 1/Cip 1>和E1 AF表达上调。瞬时表达实验证明E1 AF可以激活SiHa细胞中p21^<Waf 1/Cip 1>启动子驱动的荧光素酶报告基因。p21^<Waf 1/Cip 1>启动子活性在p53缺失的Saos 2骨肉瘤细胞中也增加，但当ets结合位点缺失时，其活性显著降低。这些结果表明，E1 AF积极调节p21^<Waf 1/Cip 1>启动子的转录，以响应遗传毒性胁迫。少

项目成果

Shindoh, M., Higashino, F. et al.: "Correlated expression of matrix netalloproteinases and ets-family transcription on factor ELA-F in imvasive oral-squamous-ull-carinoma-usived cell lins." Am. J. Pathol.148 (3). 693-700 (1996)

Shindoh, M., Higashino, F. 等人：“在侵袭性口腔鳞状细胞癌使用的细胞中，基质金属蛋白酶的相关表达与 ELA-F 因子上的 ets 家族转录相关。”

Hida, K., Shindoh, M. et al.: "Expression of E1AF,an ets-Family Transcription Factor, is Correlated with the Invasive Phenotyne of Oral Squarnvao cell Carcinomas." European Journal of Cancer (Part B), Oral Oncol.33・6. 426-430 (1997)

Hida, K., Shindoh, M. 等人：“E1AF（一种 ets-家族转录因子）的表达与口腔 Squarnvao 细胞癌的侵袭性表型相关。”《欧洲癌症杂志》（B 部分），口腔肿瘤。 33・6。426-430（1997）

Hida,K., Shindoh,M.et al.: "Antisenes E1AF Transfectlon Restrains Oral Cancer/nrasion by Reducing Matrix Metalloproteinase Activities" American Journal of Pathology. 150・6. 2125-2132 (1997)

Hida, K., Shindoh, M. 等：“Antisenes E1AF 转染通过减少基质金属蛋白酶活性抑制口腔癌/治疗”美国病理学杂志 150・6 (1997)。

Hida, k., Shindoh, M. et al.: "Antisense EIAF transfectioin restrains cral cancer cell in vasion by reducing MMPs acttrities." Am. J. Pathol.151 (in press). (1997)

Hida, k.、Shindoh, M. 等人：“反义 EIAF 转染通过减少 MMP 活性来抑制结肠癌细胞的侵袭。”

Hida, K., Shindoh, M. et al.: "Antisense E1AF Transfection Restrains Oral Cancer Invasion by Reducing Matrix Metalloproteinase Activities." American Journal of Pathology. 150・6. 2125-2132 (1997)

Hida, K., Shindoh, M. 等人：“反义 E1AF 转染通过减少基质金属蛋白酶活性抑制口腔癌侵袭。”美国病理学杂志 150・6 (1997)。