Study of the effect of Epidermal cell differentiation inhibitor on differentiation and proliferation of squamous cell carcinomas

表皮细胞分化抑制剂对鳞状细胞癌分化和增殖影响的研究

• 批准号: 08672310
• 负责人: INOUE Shingo
• 金额: $ 1.47万
• 依托单位: HIROSHIMA 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 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08672310/
• 关键词: EDIN(Epidemical cell Differentiation Inhibitor); DIFFERENTIATION; ADP-RIBOSYLATION; permeabilization

1. Effect of EDIN on the proliferation of 4 oral squamous cell carcinoma (OSCC) cell lines.The effect of EDIN was examined on the proliferation of cell lines in DMEM containing 1% or 10% FCS. HSC-2 and HSC-4 showed a little proliferative tendency in comparison with control, When 100, 250, 500 ng/ml of EDIN was added into the medium containing 1% FCS. There was no effect of EDIN on cultured cells in the medium containing 10% FCS.2. Effect of EDIN on the cell motility of OSCC cell lines.The effect of EDIN on the cell motility was investigated using Interval VTR. When 250 ng/ml of EDIN was added to the medium, H357 PRC and H357 V6 cells moved more extensively than control.3. Research for ADP-ribosylation of protein in OSCC cell lines.After maintaining in the medium with and without EDIN, cells were harvested and homogenized. ADP-ribosylation was performed with EDIN and [ィイD132ィエD1P] NAD.The autoradiograph showed that the levels of ADP-ribosylation were the same in the medium with and without EDIN in HSC-2 and HSC-4, indicating EDIN was not taken into the cells. The amount of EDIN (50, 100, 250 ng/ml) in the medium was correlated with the levels of ADP-ribosylation of 21-22 Kda protein in Fukuda cell lines, established in our department.4. Effect on the differentiation of OSCC cell lines.It was studied that EDIN might inhibit differentiation induced by CDDP using KB cell lines with permeabiization protocol of Streptolysin O. Morphological change was not observed. The levels of involucrin were measured by ELISA, but there was no difference between the presence and the absence of EDIN. Correlation wasn't recognized in the CDDP concentration and the levels of involucrin.

1. EDIN对4种口腔鳞状细胞癌细胞株增殖的影响：在含1%和10%FCS的DMEM培养基中观察EDIN对4种口腔鳞状细胞癌细胞株增殖的影响。在含1%FCS的培养液中加入100、250、500 ng/ml EDIN后，HSC-2、HSC-4的增殖趋势与对照组相比有所减弱。EDIN对含10%FCS的培养液中培养的细胞无影响. EDIN对口腔鳞癌细胞运动的影响采用Interval VTR技术观察EDIN对口腔鳞癌细胞运动的影响。当培养基中加入250 ng/ml EDIN时，H357 PRC和H357 V6细胞的运动较对照组更为广泛. OSCC细胞系中蛋白质ADP核糖基化的研究：将OSCC细胞分别置于含EDIN和不含EDIN的培养基中培养，收集细胞并匀浆。用EDIN和[腺嘌呤D132腺嘌呤D1 P] NAD进行ADP-核糖基化反应，放射自显影显示，HSC-2和HSC-4在有和无EDIN的培养基中ADP-核糖基化水平相同，表明EDIN未被摄入细胞内。EDIN的浓度（50、100、250 ng/ml）与本室建立的福田细胞系中21-22 Kda蛋白的ADP-核糖基化水平相关.对口腔鳞状细胞癌细胞系分化的影响：采用KB细胞株，用链球菌溶血素O（Streptolysin O）透化法研究EDIN对顺铂诱导的口腔鳞状细胞癌细胞系分化的抑制作用。未观察到形态学变化。ELISA法检测外表皮蛋白的含量，但EDIN的存在与不存在无差异，CDDP浓度与外表皮蛋白的含量无相关性。