Study of the inhibitory effects on malignant progression and anticancer drug sensitivity of human squamous cell carcinoma by intensifying cell to cell communication

通过加强细胞间通讯抑制人鳞状细胞癌恶性进展和抗癌药物敏感性的研究

• 批准号: 17592100
• 负责人: NAGAYASU Hiroki
• 金额: $ 1.66万
• 依托单位: Health Sciences University of Hokkaido, Institute of Personalized Medical Science
• 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-17592100/
• 关键词: oral cancer; metastasis; invasion; gap junction; anticancer drug; oral cancer; metastasis; invasion; gap junction; anticancer drug

Malatolate (Diisoprppyl, 1, 3-dithio1-2-ylidenemalonate, MT) is clinically used as a hepatoprotective agent. Because we noticed that MT induced the differetiation of cultured vascular endothelial cell, we examined the inhibitory effects of MT on invasion of oral squamous cell carcinoma cell (SCC) lines (SAS, Ca9-22, HSC-2,-3,-4) and analyzed its mechanisms of intensifying endothelial and tumor cell to cell adhesion. Five SCC cell lines and rat lung endothelial (RLE) cells were grown in a mixed medium of DMEM and D/F-12 supplemented 10% fetal bovine serum. The invasion capacity of the SCC cells was measured by counting the number of colonies /cm2 formed under the RLE monolayer using a phase contrast microscope. Intercellular junctional communication was examined by the scrape-loading assay. The percentage of cells coupled with the scrape edge was determined by the number of dye-transferred cells. Morphological changes in RLE monolayer observed with electron microscope. Connexin43 expres … More sion of RLE treated with MT was analyzed by western blotting. In an in vitro invasion assay using a RLE monolayer, invasion of tumor cells which had been treated with MT (10ng/ml, 24h) was not affected; however, when had been treated with MT, invasion was significantly inhibited in three cell lines (p<0. 01). Electron-microscopical examination of the RLE monolayer treated with MT showed the development of gap junction like structure. The cell to cell communication increased in RLE monolayers treated with MT(82. 5%), compared to non-treated with MT(32. 5%). A junction associated protein, cnnexin43 was also elevated at RLE and SAS treated with MT. In vivo growth of SAS tumor, primary tumor size was reduced in MT+CDDP treated nude mice compared with CDDP treated nude mice. Conclusion: These findings suggests that MT suppressed tumor cell invasion by intensifying the cell-cell adhesion of vascular endothelium and that enhanced anticancer drug sensitivity by modifying tumor cell communication. Less

恶性甲酸（Diisoppyl，1，3-Dithio1-2-甲状腺素，MT）在临床上用作肝保护剂。因为我们注意到MT诱导了培养的血管内皮细胞的差异，所以我们检查了MT对侵袭口服鳞状细胞癌细胞（SCC）细胞（SAS，CA9-22，HSC-2，-3，-4）的抑制作用，并分析了其强度内皮细胞和Tumor细胞的机制。在DMEM的混合培养基和D/F-12补充10％胎牛血清的混合培养基中生长了五个SCC细胞系和大鼠内皮细胞（RLE）细胞。通过计算使用相对比显微镜在RLE单层下形成的菌落 /CM2的数量来测量SCC细胞的侵袭能力。通过刮擦加载测定法检查了细胞间连通通信。结合刮擦边缘的细胞百分比由染料转让细胞的数量确定。用电子显微镜观察到的RLE单层的形态变化。 connexin43表达…通过蛋白质印迹分析了更多用MT处理的RLE。在使用RLE单层的体外侵袭测定中，未影响用MT（10ng/ml，24H）处理的肿瘤细胞的侵袭。但是，当用MT处理MT时，在三个细胞系中浸润显着抑制（p <0。01）。用MT处理的RLE单层的电子显微镜检查表明，间隙连接如结构的发展。与未经MT治疗的MT（32.5％）相比，用MT处理的RLE单层中的细胞通信增加（82.5％）。连接相关的蛋白质，CNNexin43在RLE和用MT处理的SAS上也升高。与CDDP处理过的裸鼠相比，在MT+CDDP处理的裸鼠中，MT+CDDP治疗的MT+CDDP治疗的裸小鼠的原发性肿瘤大小降低。结论：这些发现表明，MT通过增强血管内皮细胞 - 细胞粘合剂来抑制肿瘤细胞的侵袭，并通过修饰肿瘤细胞通信来增强抗癌药物敏感性。较少的