王皓,王俊杰,曲昂,等.西妥昔单抗联合照射对结直肠癌CL187细胞的抑制作用及机制探讨[J].中华放射医学与防护杂志,2012,32(5):481-484.WANG Hao,WANG Jun-jie,QU Ang,et al.Inhibitory effect of combination of cetuximab and irradiation on colorectal carcinoma CL187 cells[J].Chin J Radiol Med Prot,2012,32(5):481-484 |
西妥昔单抗联合照射对结直肠癌CL187细胞的抑制作用及机制探讨 |
Inhibitory effect of combination of cetuximab and irradiation on colorectal carcinoma CL187 cells |
投稿时间:2012-01-04 |
DOI:10.3760/cma.j.issn.0254-5098.2012.05.007 |
中文关键词: 西妥昔单抗 结直肠癌 CL187细胞 DNA损伤 |
英文关键词:Cetuximab Colorectal carcinoma CL187 cell line DNA damage |
基金项目:国家自然科学基金(81071834) |
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中文摘要: |
目的 研究西妥昔单抗(C225)联合外照射对结直肠癌CL187细胞生物学效应的影响,并探讨相关分子机制。方法 结直肠癌CL187细胞分单纯照射组和C225处理的联合照射组,受6 MV X 射线照射0、4和8 Gy后24和48 h,用四甲基偶氮唑盐(MTT)比色法测吸光度(A)值,比较两组细胞死亡率的差异。利用克隆形成实验比较两组细胞增殖能力的差异。采用流式细胞仪,检测细胞周期和细胞凋亡。蛋白免疫印迹法分析两组细胞DNA-PKcs、Ku70和Ku80蛋白表达量的变化。结果 联合照射组较单纯照射组死亡细胞比例增加(t=-6.14、-6.53,P<0.05),细胞克隆形成能力下降。C225增强了射线对细胞的杀伤作用,放射增敏比SER为1.38。联合照射组G0/G1期细胞阻滞增加(t=-4.64, P<0.05),细胞凋亡比例增加(t=-9.16, P<0.05),DNA修复相关蛋白DNA-PKcs、Ku70和Ku80蛋白表达量减少。结论 西妥昔单抗增强照射对结直肠癌CL187细胞的杀伤作用,可能是通过影响细胞周期、细胞凋亡和DNA损伤修复基因实现的。 |
英文摘要: |
Objective To investigate the combination effect of cetuximab and irradiation on colorectal carcinoma CL187 cell line and underlying molecular mechanism.Methods CL187 cells with or without cetuximab treatment were irradiated by 0, 4 and 8 Gy X-rays, then cell death percentage was determined by MTT 24 and 48 h post-irradiation. Clone forming assay was used to evaluate the cell reproliferation ability. Cell cycle distribution, apoptosis, and necrosis were analyzed by flow cytometry. Western blot was used to detect the protein expressions of DNA-PKcs, Ku70 and Ku80.Results The cetuximab enhanced the percentage of radiation-induced cell death, while descreased the cloning formation capacity and increased radiosenvtivity (t=-6.14、-6.53, P<0.05). The SER of cetuximab on CL187 cell line approached to 1.38. In addition, cetuximab also increased radiation-induced G0/G1 phase arrest (t=-4.64, P<0.05) and the percentage of apoptosis and necrosis (t=-9.16, P<0.05), but it descreased the expression levels of DNA-PKcs, Ku70 and Ku80 proteins. Conclusions The cetuximab treatment might enhance the inhibitory effect of irradiation on colorectal carcinoma CL187 cell line by influencing cell cycle distribution, cell apoptosis, and the expression of DNA repair proteins. |
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